Journal: International Journal of Molecular Sciences
Article Title: TAS1R3 Regulates GTPase Signaling in Human Skeletal Muscle Cells for Glucose Uptake
doi: 10.3390/ijms27010103
Figure Lengend Snippet: Lactisole inhibits insulin-stimulated glucose uptake and Rac1 activation in human LHCN-M2 myotubes. ( A , B ) Human LHCN-M2 myotubes were pre-treated with vehicle (DMSO) or inhibitor for 1 h and then stimulated with ± 100 nM insulin 5 minor for 20 min for Rac1 activation and glucose uptake assay, respectively. TAS1R3(i); lactisole. Gαq/11(i); YM-25490. Assessment of [ 3 H]2-DG uptake ( A ) and Rac1 activation ( B ). ( A ) Glucose uptake was normalized to the protein content. Cpm: Counts per minute ( n = 4 independent experiments). ( B ) Rac1 activation values are relative to baseline Rac1-GTP levels (set to 1.0) detected with unstimulated basal insulin ( n = 5 independent experiments). ( C , D ) Human LHCN-M2 myotubes were pre-treated with siRNA (48 h) and then stimulated with ±100 nM insulin (10 min). CON: control. ( C ) Impact of TAS1R3 siRNA knockdown. Top: Representative immunoblot (IB). Bottom: Densitometry analysis of TAS1R3 protein expression normalized to tubulin (Tub). Control-siRNA values were set to 1.0. n = 4 independent experiments. ( D ) Effects of siRNA treatment on glucose uptake with insulin stimulation ( n = 8 independent experiments). Glucose uptake was normalized to the protein content. Cpm: Counts per minute. Values under basal unstimulated insulin conditions were set to 1.0. ( A – D ) Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. N.S: not significant.
Article Snippet: Rac1 activation was measured in the supernatant using the commercially available G-LISA Rac1 Activation Assay from Cytoskeleton Inc. (Cat # BK 128), as described previously [ ].
Techniques: Activation Assay, Control, Knockdown, Western Blot, Expressing
Cytoskeleton Inc is a verified supplier 
Cytoskeleton Inc manufactures this product 
![Lactisole inhibits insulin-stimulated glucose uptake and <t>Rac1</t> activation in human LHCN-M2 myotubes. ( A , B ) Human LHCN-M2 myotubes were pre-treated with vehicle (DMSO) or inhibitor for 1 h and then stimulated with ± 100 nM insulin 5 minor for 20 min for Rac1 activation and glucose uptake assay, respectively. TAS1R3(i); lactisole. Gαq/11(i); YM-25490. Assessment of [ 3 H]2-DG uptake ( A ) and Rac1 activation ( B ). ( A ) Glucose uptake was normalized to the protein content. Cpm: Counts per minute ( n = 4 independent experiments). ( B ) Rac1 activation values are relative to baseline Rac1-GTP levels (set to 1.0) detected with unstimulated basal insulin ( n = 5 independent experiments). ( C , D ) Human LHCN-M2 myotubes were pre-treated with siRNA (48 h) and then stimulated with ±100 nM insulin (10 min). CON: control. ( C ) Impact of TAS1R3 siRNA knockdown. Top: Representative immunoblot (IB). Bottom: Densitometry analysis of TAS1R3 protein expression normalized to tubulin (Tub). Control-siRNA values were set to 1.0. n = 4 independent experiments. ( D ) Effects of siRNA treatment on glucose uptake with insulin stimulation ( n = 8 independent experiments). Glucose uptake was normalized to the protein content. Cpm: Counts per minute. Values under basal unstimulated insulin conditions were set to 1.0. ( A – D ) Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. N.S: not significant.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_6175/pmc12786175/pmc12786175__ijms-27-00103-g002.jpg)
