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. 2019 Sep 12;294(44):16172–16185. doi: 10.1074/jbc.RA119.009110

Figure 7.

Figure 7.

KLHL9/KLHL13/CUL3 E3 Ub ligase complex regulates IRS1 protein content and insulin action in ATG16L1 KO MEFs. A, Klhl9; B, Klhl13; and C, Cul3 siRNA-transfected WT and ATG16L1 KO MEFs (48 h) were examined for IRS1 expression (n = 3–4). p value was calculated using a two-tailed unpaired t test. D, Klhl9; E, Klhl13; and F, Cul3 siRNA-transfected WT and ATG16L1 KO MEFs were treated with insulin (10 nm, 10 min) and immunoblotted for pAKT on Ser-473 (n = 3–4). p value was calculated using a two-way ANOVA with Tukey's post hoc. siRNA knockdown efficiency: Klhl9 WT, 90.6 ± 2.3%; ATG16L1 KO, 84.7 ± 4.8%; Klhl13 WT, 96.7 ± 0.8%; ATG16L1 KO, 91.4 ± 2.5%; and Cul3 WT, 70.4 ± 2.0%; ATG16L1 KO, 66.9 ± 6.5%. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.