Figure 3. Effect of p62 knockout and the mutation in the LC3-interacting region motif of p62 in embryonic stem cells and neurons.

(A) mScarlet–p62 WT and mScarlet–p62 LC3-interacting region mutant embryonic stem cells were treated for 3 h with either 400 nM bafilomycin or 10 μM MG-132 or a combination of the two drugs. A DMSO-treated sample served as a negative control. Western blot analysis was performed using the antibodies indicated. (A, B) Quantification of the Western blot data from (A). p62 and NBR1 band intensity values were normalised to the GAPDH-loading control. The graphs represent the mean of the GAPDH-normalised values ± SEM from n = 3 independent experiments. P-values were calculated with a two-way ANOVA and Tukey’s multiple comparison test (P > 0.05 non-significant [not shown], 0.01 < P < 0.05 *, 0.001 < P < 0.01 **, P < 0.001 ***). (C) DIV21 neurons were treated for 5 h with 400 nM bafilomycin. Cell lysates were analysed by Western blot.