phospho insr  (Cell Signaling Technology Inc)

Journal: Cell Death & Disease

Article Title: FAM3A drives uncoupling of muscle lipid accumulation and insulin resistance depending on insulin receptor

doi: 10.1038/s41419-025-08298-1

Figure Lengend Snippet: a Western blot images showing the presence of FAM3A in different cellular compartments in soleus muscles and heart tissues. b Levels of the FAM3A protein along with positive controls (TSG101 and CD9) and the negative control (calnexin) in plasma-derived exosomes from patients chosen at random. c Western blot images and quantification of the levels of p-AKT (thr308) and p-insR-β (tyr1150/1151) in cells treated with different concentrations (ng/ml) of rcFAM3A for 8 h (n = 4 biologically independent samples/group; quantitative comparisons between samples were run on the same gel). d Western blot images and quantification of the expression levels of the indicated proteins in soleus muscles from the mice treated with bms536924 and fed an HFD for 10 weeks (n = 6 biologically independent animals/group; quantitative comparisons between samples were run on the same gel). e Western blot images and quantification of the expression levels of the indicated proteins in cells pre-treated with bms536924 or transfected with the insR siRNA and then stimulated with rcFAM3A (200 ng/ml; n = 4 biologically independent samples/group; quantitative comparisons between samples were run on the same gel). The lipid droplets detected by oil red O staining ( f ) and TG content ( g ) were measured in soleus muscles from the mice treated with bms536924 and fed an HFD for five weeks and the results were plotted (n = 6 biologically independent animals/group). Scale bar: 200 μm, insets: 100 μm in ( f ). C2C12 cells were treated with rcFAM3A (200 ng/ml) for 12 hours. Cellular De novo FA synthesis ( h ) and TG content ( i ) were measured and graphed (n = 3 biologically independent samples/group). j , k Body adiposity and body weight gain were measured in mice treated with bms536924 and fed an HFD for 10 weeks and the results were plotted (n = 7 biologically independent animals/group). The data are presented as the means ± SEMs. Statistical significance was determined with a two-tailed independent t test, and P values are indicated ( ns P ≥ 0.05). Source data are provided as a file.

Article Snippet: Anti-FAM3A (Origene, #TA324017; 1:1000), anti-adiponectin (Cell Signaling Technology, #2789; 1:1000), anti-PPARα (Santa Cruz, #sc-398394; 1:500), anti-srebp1 (Santa Cruz, #sc-13551; 1:500), anti-fas (Cell Signaling Technology, #3180; 1:1000), anti-acly (Cell Signaling Technology, #4332; 1:1000), anti-acc (Cell Signaling Technology, #3676; 1:1000), anti-p65 (Santa Cruz, #sc-8008; 1:500), anti-pcna (Santa Cruz, #sc-25280; 1:500), anti-p-AKT (thr308) (Cell Signaling Technology, #9275; 1:1000), anti-AKT (Cell Signaling Technology, #9272; 1:1000), anti-p-insR-β (tyr1150/1151) (Cell Signaling Technology, #3024; 1:1000), anti-insR-β (Cell Signaling Technology, #3025; 1:1000), anti-Calnexin (Abcam, #ab75801; 1:1000), anti-TSG101 (Abcam, #ab125011; 1:1000), anti-CD9 (Abcam, #ab92726; 1:1000), anti-β-actin (LABLEAD, #A0101; 1:4000), anti-GAPDH (Cell Signaling Technology, #5174; 1:4000), and HRP-conjugated secondary antibodies were commercially obtained (Supplementary Table ).

Techniques: Western Blot, Muscles, Negative Control, Clinical Proteomics, Derivative Assay, Expressing, Transfection, Staining, Two Tailed Test