Ular compartment in the basal state [25]. Furthermore, GLUT1 may also influence

Ular compartment in the basal state [25]. Furthermore, GLUT1 may also influence and cooperate with GLUT12. In this study, GLUT12 expression influenced the transcription of GLUT1, which 10781694 was responsible for the basal Title Loaded From File glucose uptake of the cells. This decreased GLUT1 expression and increased GLUT12 expression in the GT12-GMGE cells implies that GLUT12 may provide a back-up system for GLUT1, inducing an increase in glucose consumption at 48 h. Given that GLUT12 is distributed in the cytoplasm and GLUT1 is responsible for the absorption of glucose from the basal plasma membrane of epithelium, this may be related to lactose synthesis. Solute carrier family 1 member 5 (SLC1A5) encodes ASCT2, the major amino acid transporter, which transports the neutralFunctional Analysis of GLUT1 and GLUTFigure 4. Immunofluorescence localization of goat GLUT1 and GLUT12 in GT1-GMGE and GT12-GMGE. Cells were fixed with paraformaldehyde, permeablized and incubated with antibody of GLUT1 or GLUT12. At last, the plate incubated with DAPI. The green fluorescence was scattered Title Loaded From File throughout the cytoplasm in GMGE. In GT1-GMGE cells, the total GLUT1 was mainly distributed along the plasma membrane in addition to the cytoplasm. The red fluorescence was mainly distributed around the nuclear in GMGE, while the GLUT12 expression was increased and was also mainly distributed in the perinuclear region in GT12-GMGE cells. Original magnification 4006. doi:10.1371/journal.pone.0065013.gamino acids Ala, Gly, Leu, Ile, Val, Ser, Cys, Thr and Pro. The mRNA expression of SLC1A5 in the GT1-GMGE and GT12GMGE cells was significantly increased compared to the GMGE cells, indicating that protein synthesis and metabolism in the GT1GMGE and GT12-GMGE cells were higher than in the GMGE cells, processes that are related to increased glucose uptake [26]. We also found that the mRNA levels of SLC7A5 and SLC3A2 were also increased. Solute carrier family 7 member 5 (SLC7A5) codes for LAT1, which transports cationic amino acids; solute carrier family 3 member 2 (SLC3A2) codes for 4F2hc, which is an activator of dibasic and neutral amino acid transporters. Two AATs form the functional heterodimer AA transporter and have a reciprocal regulatory connection with mTOR in the regulation of protein synthesis and cell viability, most likely through the active transport of Leu [27]. mTOR controls ribosome biogenesis and cell growth in regulatory pathways. New Title Loaded From File research shows that the mTOR Title Loaded From File pathway is influenced by the intracellular concentration of ATP, independent of the abundance of amino acids, and that mTOR itself is an ATP sensor [26]. Thus, the additional glucose due to the overexpression of GLUT1 may promote protein synthesis and strengthen the cell viability. 4F2hc is a membraneprotein that interacts with GLUT1 and is likely to be involved in GLUT1 stabilization and contributes to the regulation of glucose metabolism [28]. The results of our study corroborate the findings of Ohno et al. who reported GLUT1 functions require the assistance of 4F2hc [28]. Many proteins are targeted to the plasma membrane, including human GLUT4 and GLUT12, and are transported from the endoplasmic reticulum through the Golgi apparatus to the cell surface [4,29]. Human GLUT4 and GLUT12 contain several potential targeting motifs that control their localization. Dileucine motifs are present in both the NH2 and COOH termini in human GLUT12. In the absence of insulin, GLUT12 is localized to a perinuclear site in MCF-7 cells [.Ular compartment in the basal state [25]. Furthermore, GLUT1 may also influence and cooperate with GLUT12. In this study, GLUT12 expression influenced the transcription of GLUT1, which 10781694 was responsible for the basal glucose uptake of the cells. This decreased GLUT1 expression and increased GLUT12 expression in the GT12-GMGE cells implies that GLUT12 may provide a back-up system for GLUT1, inducing an increase in glucose consumption at 48 h. Given that GLUT12 is distributed in the cytoplasm and GLUT1 is responsible for the absorption of glucose from the basal plasma membrane of epithelium, this may be related to lactose synthesis. Solute carrier family 1 member 5 (SLC1A5) encodes ASCT2, the major amino acid transporter, which transports the neutralFunctional Analysis of GLUT1 and GLUTFigure 4. Immunofluorescence localization of goat GLUT1 and GLUT12 in GT1-GMGE and GT12-GMGE. Cells were fixed with paraformaldehyde, permeablized and incubated with antibody of GLUT1 or GLUT12. At last, the plate incubated with DAPI. The green fluorescence was scattered throughout the cytoplasm in GMGE. In GT1-GMGE cells, the total GLUT1 was mainly distributed along the plasma membrane in addition to the cytoplasm. The red fluorescence was mainly distributed around the nuclear in GMGE, while the GLUT12 expression was increased and was also mainly distributed in the perinuclear region in GT12-GMGE cells. Original magnification 4006. doi:10.1371/journal.pone.0065013.gamino acids Ala, Gly, Leu, Ile, Val, Ser, Cys, Thr and Pro. The mRNA expression of SLC1A5 in the GT1-GMGE and GT12GMGE cells was significantly increased compared to the GMGE cells, indicating that protein synthesis and metabolism in the GT1GMGE and GT12-GMGE cells were higher than in the GMGE cells, processes that are related to increased glucose uptake [26]. We also found that the mRNA levels of SLC7A5 and SLC3A2 were also increased. Solute carrier family 7 member 5 (SLC7A5) codes for LAT1, which transports cationic amino acids; solute carrier family 3 member 2 (SLC3A2) codes for 4F2hc, which is an activator of dibasic and neutral amino acid transporters. Two AATs form the functional heterodimer AA transporter and have a reciprocal regulatory connection with mTOR in the regulation of protein synthesis and cell viability, most likely through the active transport of Leu [27]. mTOR controls ribosome biogenesis and cell growth in regulatory pathways. New research shows that the mTOR pathway is influenced by the intracellular concentration of ATP, independent of the abundance of amino acids, and that mTOR itself is an ATP sensor [26]. Thus, the additional glucose due to the overexpression of GLUT1 may promote protein synthesis and strengthen the cell viability. 4F2hc is a membraneprotein that interacts with GLUT1 and is likely to be involved in GLUT1 stabilization and contributes to the regulation of glucose metabolism [28]. The results of our study corroborate the findings of Ohno et al. who reported GLUT1 functions require the assistance of 4F2hc [28]. Many proteins are targeted to the plasma membrane, including human GLUT4 and GLUT12, and are transported from the endoplasmic reticulum through the Golgi apparatus to the cell surface [4,29]. Human GLUT4 and GLUT12 contain several potential targeting motifs that control their localization. Dileucine motifs are present in both the NH2 and COOH termini in human GLUT12. In the absence of insulin, GLUT12 is localized to a perinuclear site in MCF-7 cells [.Ular compartment in the basal state [25]. Furthermore, GLUT1 may also influence and cooperate with GLUT12. In this study, GLUT12 expression influenced the transcription of GLUT1, which 10781694 was responsible for the basal glucose uptake of the cells. This decreased GLUT1 expression and increased GLUT12 expression in the GT12-GMGE cells implies that GLUT12 may provide a back-up system for GLUT1, inducing an increase in glucose consumption at 48 h. Given that GLUT12 is distributed in the cytoplasm and GLUT1 is responsible for the absorption of glucose from the basal plasma membrane of epithelium, this may be related to lactose synthesis. Solute carrier family 1 member 5 (SLC1A5) encodes ASCT2, the major amino acid transporter, which transports the neutralFunctional Analysis of GLUT1 and GLUTFigure 4. Immunofluorescence localization of goat GLUT1 and GLUT12 in GT1-GMGE and GT12-GMGE. Cells were fixed with paraformaldehyde, permeablized and incubated with antibody of GLUT1 or GLUT12. At last, the plate incubated with DAPI. The green fluorescence was scattered throughout the cytoplasm in GMGE. In GT1-GMGE cells, the total GLUT1 was mainly distributed along the plasma membrane in addition to the cytoplasm. The red fluorescence was mainly distributed around the nuclear in GMGE, while the GLUT12 expression was increased and was also mainly distributed in the perinuclear region in GT12-GMGE cells. Original magnification 4006. doi:10.1371/journal.pone.0065013.gamino acids Ala, Gly, Leu, Ile, Val, Ser, Cys, Thr and Pro. The mRNA expression of SLC1A5 in the GT1-GMGE and GT12GMGE cells was significantly increased compared to the GMGE cells, indicating that protein synthesis and metabolism in the GT1GMGE and GT12-GMGE cells were higher than in the GMGE cells, processes that are related to increased glucose uptake [26]. We also found that the mRNA levels of SLC7A5 and SLC3A2 were also increased. Solute carrier family 7 member 5 (SLC7A5) codes for LAT1, which transports cationic amino acids; solute carrier family 3 member 2 (SLC3A2) codes for 4F2hc, which is an activator of dibasic and neutral amino acid transporters. Two AATs form the functional heterodimer AA transporter and have a reciprocal regulatory connection with mTOR in the regulation of protein synthesis and cell viability, most likely through the active transport of Leu [27]. mTOR controls ribosome biogenesis and cell growth in regulatory pathways. New research shows that the mTOR pathway is influenced by the intracellular concentration of ATP, independent of the abundance of amino acids, and that mTOR itself is an ATP sensor [26]. Thus, the additional glucose due to the overexpression of GLUT1 may promote protein synthesis and strengthen the cell viability. 4F2hc is a membraneprotein that interacts with GLUT1 and is likely to be involved in GLUT1 stabilization and contributes to the regulation of glucose metabolism [28]. The results of our study corroborate the findings of Ohno et al. who reported GLUT1 functions require the assistance of 4F2hc [28]. Many proteins are targeted to the plasma membrane, including human GLUT4 and GLUT12, and are transported from the endoplasmic reticulum through the Golgi apparatus to the cell surface [4,29]. Human GLUT4 and GLUT12 contain several potential targeting motifs that control their localization. Dileucine motifs are present in both the NH2 and COOH termini in human GLUT12. In the absence of insulin, GLUT12 is localized to a perinuclear site in MCF-7 cells [.Ular compartment in the basal state [25]. Furthermore, GLUT1 may also influence and cooperate with GLUT12. In this study, GLUT12 expression influenced the transcription of GLUT1, which 10781694 was responsible for the basal glucose uptake of the cells. This decreased GLUT1 expression and increased GLUT12 expression in the GT12-GMGE cells implies that GLUT12 may provide a back-up system for GLUT1, inducing an increase in glucose consumption at 48 h. Given that GLUT12 is distributed in the cytoplasm and GLUT1 is responsible for the absorption of glucose from the basal plasma membrane of epithelium, this may be related to lactose synthesis. Solute carrier family 1 member 5 (SLC1A5) encodes ASCT2, the major amino acid transporter, which transports the neutralFunctional Analysis of GLUT1 and GLUTFigure 4. Immunofluorescence localization of goat GLUT1 and GLUT12 in GT1-GMGE and GT12-GMGE. Cells were fixed with paraformaldehyde, permeablized and incubated with antibody of GLUT1 or GLUT12. At last, the plate incubated with DAPI. The green fluorescence was scattered throughout the cytoplasm in GMGE. In GT1-GMGE cells, the total GLUT1 was mainly distributed along the plasma membrane in addition to the cytoplasm. The red fluorescence was mainly distributed around the nuclear in GMGE, while the GLUT12 expression was increased and was also mainly distributed in the perinuclear region in GT12-GMGE cells. Original magnification 4006. doi:10.1371/journal.pone.0065013.gamino acids Ala, Gly, Leu, Ile, Val, Ser, Cys, Thr and Pro. The mRNA expression of SLC1A5 in the GT1-GMGE and GT12GMGE cells was significantly increased compared to the GMGE cells, indicating that protein synthesis and metabolism in the GT1GMGE and GT12-GMGE cells were higher than in the GMGE cells, processes that are related to increased glucose uptake [26]. We also found that the mRNA levels of SLC7A5 and SLC3A2 were also increased. Solute carrier family 7 member 5 (SLC7A5) codes for LAT1, which transports cationic amino acids; solute carrier family 3 member 2 (SLC3A2) codes for 4F2hc, which is an activator of dibasic and neutral amino acid transporters. Two AATs form the functional heterodimer AA transporter and have a reciprocal regulatory connection with mTOR in the regulation of protein synthesis and cell viability, most likely through the active transport of Leu [27]. mTOR controls ribosome biogenesis and cell growth in regulatory pathways. New research shows that the mTOR pathway is influenced by the intracellular concentration of ATP, independent of the abundance of amino acids, and that mTOR itself is an ATP sensor [26]. Thus, the additional glucose due to the overexpression of GLUT1 may promote protein synthesis and strengthen the cell viability. 4F2hc is a membraneprotein that interacts with GLUT1 and is likely to be involved in GLUT1 stabilization and contributes to the regulation of glucose metabolism [28]. The results of our study corroborate the findings of Ohno et al. who reported GLUT1 functions require the assistance of 4F2hc [28]. Many proteins are targeted to the plasma membrane, including human GLUT4 and GLUT12, and are transported from the endoplasmic reticulum through the Golgi apparatus to the cell surface [4,29]. Human GLUT4 and GLUT12 contain several potential targeting motifs that control their localization. Dileucine motifs are present in both the NH2 and COOH termini in human GLUT12. In the absence of insulin, GLUT12 is localized to a perinuclear site in MCF-7 cells [.