Figure 6.

CIP2A is a CMA substrate. (A) Immunoblot for CIP2A and ACTB protein in total protein lysates of NIH-3T3 cells, control (ctrl) or LAMP2A knocked down (LAMP2A [-]), transduced with a plasmid encoding MYC or the empty vector. (B) Densitometric quantification for CIP2A in blots as the one shown in (A). Values are relative to ctrl empty vector cells and normalized to Ponceau S staining (N = 6). ((C)and D) NIH-3T3 MYC-transduced cells, ctrl or LAMP2A (-) 12 h after incubation without additions (None) or in the presence of lysosomal proteolysis inhibitors (ammonium chloride and leupeptin, N/L) or a proteasome inhibitor (MG132). (C) Immunoblot for the indicated proteins. (D) Densitometry analysis of CIP2A in blots as the one shown in C. Values are relative to None condition and normalized to Ponceau S staining (N = 6). In both graphs values are presented as mean ± SEM. Two-way ANOVA and the Bonferroni post-hoc test were used and differences were considered significant for *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001. ns represents nonsignificant changes.