Fig. 5: Gab2 deficiency increases the expression of thermogenesis-related genes in HFD-fed mice and differentiated brown adipocytes. | Cell Death & Disease

Fig. 5: Gab2 deficiency increases the expression of thermogenesis-related genes in HFD-fed mice and differentiated brown adipocytes.

From: Gab2 deficiency suppresses high-fat diet-induced obesity by reducing adipose tissue inflammation and increasing brown adipose function in mice

Fig. 5

A Representative IHC staining images of Ucp1 in BAT from Gab2 KO and WT mice fed with HFD diet for 12 weeks. The scale bar is 50 μm. B Left panel, representative blots of Ucp1 and Gab2 protein expressions in BAT from Gab2 KO and WT mice fed with HFD diet for 12 weeks detected by immunoblotting analysis. Middle and right panel, the relative grayscale values of Ucp1 (middle) and Gab2 (right) protein blots were normalized with the loading control protein β-Tubulin. C, D The mRNA level of thermogenesis-related genes (C) and mitochondrial function associated genes (D) in BAT from Gab2 KO and WT mice fed with HFD diet for 12 weeks detected by RT-qPCR. The results were normalized with S18 mRNA levels. E Oil Red O staining of completely differentiated brown adipocytes (Day 6). F Left panel, representative blots of Ucp1 and Gab2 protein expressions during the differentiation of brown preadipocytes detected by immunoblotting analysis. Middle and right panel, the relative grayscale values of Ucp1 (middle) and Gab2 (right) protein blots were normalized with the loading control protein β-Tubulin. G The mRNA level of Gab2 (left panel) and Ucp1 (right panel) during the differentiation of brown preadipocytes detected by RT-qPCR. The results were normalized with S18 mRNA levels. H Ucp1-luciferase reporter activity in response to Gab2 overexpression (left panel) or Gab2 knockdown (right panel) analyzed in immortalized brown preadipocytes by using the Dual-Luciferase Reporter Assay System (Promega). Con, immortalized brown preadipocytes only transfected with Ucp1-luciferase reporter and renilla; si-con, siRNA of control protein. I Representative blots of Ucp1 and Gab2 protein expressions in undifferentiated and differentiated brown adipocytes detected by immunoblotting analysis. β-Tubulin protein was measured as a loading control. Con, undifferentiated brown preadipocytes; Induced, differentiated brown adipocytes. J The mRNA level of common or brown fat-selective genes in differentiated brown adipocytes (Day 6) detected by RT-qPCR. The results were normalized with S18 mRNA levels. The data are presented as mean ± SD from at least six mice in each group or three independent experiments. Statistical difference is indicated: *P < 0.05; **P < 0.01; ***P < 0.001.

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