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. 2020 Dec 8;134(1):73–80. doi: 10.1097/CM9.0000000000001255

Figure 3.

Figure 3

GAB1 natively regulates autophagy. (A) HUVECs were treated with either control siRNA (siControl) or GAB1-targeting siRNA (siGAB1) for 48 h, followed by treatment of HBSS for 6 h for the induction of autophagy presented with or without BAF. Cell lysates were harvested to examine the protein levels of GAB1 (using anti-GAB1 antibody), LC3I/II, and β-actin by Western blotting. (B) HeLa cells were transiently transfected with control or GAB1 expression vector control for 24 h, followed by the treatment as mentioned above. Western blotting was performed and analyzed as mentioned above. (C) Protein levels of LC3II were quantitated by densitometric analysis using Image J normalized to β-actin and presented as fold changes compared with the control group. ∗siControl vs. siGAB1 in HBSS− and BAF−; †siControl vs. siGAB1 in HBSS+ and BAF−; ‡siControl vs. siGAB1 in HBSS+ and BAF+. (D) Protein levels of LC3II were quantitated by densitometric analysis using Image J normalized to β-actin and presented as fold changes compared with the control group. ∗Control vs. GAB1 in HBSS− and BAF−; †Control vs. GAB1 in HBSS+ and BAF−; ‡Control vs. GAB1 in HBSS+ and BAF+. ASO: Arteriosclerosis obliterans; BAF: Bafilomycin A1; DAPI: 4′,6-Diamidino-2-phenylindole; GAB1: GRB2 associated binding protein 1; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; HBSS: Hank balanced salt solution; HUVEC: Human umbilical vein endothelial cells.