Fig. 5: PPTC7 is an essential co-factor of SCFFBXL4 E3 ubiquitin ligase complex. | Cell Death & Disease

Fig. 5: PPTC7 is an essential co-factor of SCFFBXL4 E3 ubiquitin ligase complex.

From: PPTC7 acts as an essential co-factor of the SCFFBXL4 ubiquitin ligase complex to restrict BNIP3/3L-dependent mitophagy

Fig. 5

A WB analysis of the indicated proteins in the input from parental HeLa cells stably overexpressing EV, PPTC7-WT-FLAG, or PPTC7-D78A-FLAG mutant. B RT-qPCR measurement of BNIP3 and BNIP3L mRNA levels in HeLa cells stably overexpressing EV or PPTC7-FLAG. Data are shown as means ± SD (n = 3). C WB analysis of the indicated proteins in WCL from FBXL4 KO HeLa cells stably overexpressing EV or PPTC7-FLAG. D, E Representative IF images from FBXL4 KO HeLa cells stably overexpressing EV, PPTC7-WT-FLAG, or PPTC7-D78A-FLAG mutant, stained with BNIP3 (or BNIP3L), HSP60, and DAPI. Scale bar, 10 μm. The relative intensity of HSP60, BNIP3, and BNIP3L was quantified and shown in (E). Data were shown as means ± SD (n = 20 cells per group). F WB analysis of the indicated proteins in WCL from PPTC7 KO HeLa cells stably overexpressing EV or FLAG-FBXL4. G, H WB of the products of in vivo ubiquitination assays from parental HeLa cells transfected with the indicated plasmids. I, J WB of the products of in vivo ubiquitination assays from PPTC7 KO HeLa cells transfected with the indicated plasmids. P values are calculated by the Two-way ANOVA test in (B, E). **P < 0.01.

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