Fig. 3: DNAJA2 regulates PCM1/CEP290 degradation via chaperone-mediated autophagy.

a Western blotting analysis showing PCM1 and CEP290 protein levels in 4T1 cells treated with 20 μM chloroquine (CQ) for the indicated times. b Quantifications of relative protein levels (n = 4) of PCM1 and CEP290 as shown in (a). c Western blotting analysis showing PCM1 protein level in WT and DJ2^−/− HeLa cells treated with or without NL (20 mM ammonium chloride and 100 μM leupeptin) for 16 h. d Calculation of PCM1 degradation rate (n = 3) in lysosomes in WT and DJ2^−/− HeLa cells as shown in (c). Lysosomal degradation rate was calculated as: degradation per hour (%) = 100%(Relative PCM1 level of NL-treated group/Relative PCM1 level of NL-untreated group - 1)/16. e Western blotting analysis showing PCM1 and CEP290 protein levels in HeLa cells treated with 10 μM Apoptozole (AZ) for the indicated times. f Quantifications of relative protein levels (n = 4) of PCM1 and CEP290 as shown in (e). g Western blotting analysis showing the expression levels of PCM1 and CEP290 in WT and L2A^−/− 4T1 cells. * indicates a non-specific band. h Quantifications of relative PCM1 and CEP290 levels (n = 3) in WT and L2A^−/− 4T1 cells, as shown in (g). i Protein stability analysis of PCM1 and CEP290 in WT and L2A^−/− 4T1 cells treated with 50 μg/ml cycloheximide (CHX), as indicated. j, k Quantifications of relative protein levels (n = 3) of PCM1 and CEP290 as shown in (i). l Western blotting analysis of PCM1 degradation in response to AR7 (a CMA activator) in WT, DJ2^−/−, DJ2-rescued and L2A^−/− 4T1 cells. m Calculation of PCM1 degradation rate (n = 4) in response to AR7 treatment in WT, DJ2^−/−, DJ2-rescued and L2A^−/− 4T1 cells as shown in (l). Data are shown as means ± SEM of n experimental repeats. The P values in figures (b), (f) were determined by two-tailed unpaired t test and the others were determined by two-tailed unpaired t test with Welch’s correction. Source data are provided as a Source data file.