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. 2023 Jan 13;32(10):1660–1672. doi: 10.1093/hmg/ddad006

Figure 6.

Figure 6

The core p62-binding domain in SSH1 is required to suppress autophagy and mitophagy. (A) Representative images of HT22 cells co-expressing the respective Flag tagged SSH1 constructs (blue) and mCherry-GFP-p62 (red and green), with or without 5 μm FCCP treatment for 4 h. (B, C) Quantification of the percentage of red p62 puncta (acidified) over the total amount of p62 puncta for control condition of A. Data are presented as mean ± SEM. n = 10–12 images/condition from three independent experiments, one-way ANOVA, followed by Dunnett’s post hoc, #P < 0.0001, ***P < 0.0005, *P < 0.05. (B, D) Quantification of the percentage of red p62 puncta (acidified) over the total amount of p62 puncta for FCCP condition of A. Data are presented as mean ± SEM. n = 10–12 images/condition from three independent experiments, one-way ANOVA, followed by Dunnett’s post hoc, #P < 0.0001. (E) Representative images of HT22 cells co-expressing mito-QC (mCherry-GFP-Fis1) (red and green) and VC, Flag-SSH1, Flag-SSH1ΔN-ΔC649 or Flag-SSH1ΔN-ΔC549 (blue), with or without 5 μm FCCP treatment for 4 h. (F) Quantification of red (acidified) mito-QC puncta for E. Data are presented as mean ± SEM. n = 8–10 images/condition from four independent experiments, one-way ANOVA, followed by Dunnett’s post hoc, #P < 0.0001.