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. 2022 Jul 21;13:4220. doi: 10.1038/s41467-022-31869-1

Fig. 1. CA39 and CA77 activate CMA in a dose-dependent manner.

Fig. 1

a Chemical structure of AR7, CA39, and CA77. b Molecular docking of CA39 (orange sticks) and CA77 (green sticks) in the binding pocket of inactive RARα (PDB ID: 1DKF). c Interaction map showing predicted RARα amino acid interactions with CA39 and CA77. d CMA activity in NIH3T3 stably expressing the KFERQ-PS-Dendra after incubation with AR7, CA39, or CA77. Representative images of cells treated with 20 μM of each compound. Nuclei are highlighted with DAPI. Inserts shows higher magnification of the red channel. Right shows the comparison with activation of CMA by AR7. n = 3 independent experiments (>1,500 cells counted). e–g Quantification of CMA activity in the same cells upon addition of increasing concentrations of CA39 (e), CA77 (f) or AR7 (g) for 12 (left) or 24 (right) hours. n = 4 independent experiments (>2,500 cells counted). h–j Quantification of CMA activity in the same cells after addition of increasing concentrations of CA39 (h), CA77 (i) or AR7 (j) for 12, 24 h, and 12 h after washing (w) them out from the media. n = 3 independent experiments (>1,500 cells counted). All values are mean + s.e.m. One-way ANOVA (d–g) or two-way ANOVA (h, j) followed by Bonferroni’s multiple comparisons post-hoc test were used. Significant differences with untreated samples are indicated in e and among the different incubation protocols in f. **p < 0.01, ***p < 0.001, ****p < 0.0001. ns: not significant. Source data and exact p values are provided as a Source Data file.