Figure 5.
PAN3 activates pexophagy. (A) PAN3 is represented as a prominent teal dot on the volcano plot. Red dots on the graph denote common regulators across all four conditions. (B) Control and PAN3 KO HEK293A cells were treated with clofibrate (1 mM) for 6 h in the presence or absence of Baf. Changes in p62 and LC3B levels were analyzed using WB. (C) Control and KO cells were incubated with clofibrate (1 mM) for 6 h. p62 and LC3B puncta were visualized and quantified by IF. Scale bars, 10 µM. (D) Control and KO cells were treated with clofibrate in the presence or absence of Baf. WB was then used to examine the pexophagy receptor, PMP70. (E) Indicated cells were incubated with clofibrate. PMP70 signal was visualized and quantified by IF. Scale bars, 10 µM. (F) Control and PAN3 KD HCT116 cells stably expressing the RFP-GFP-SKL reporter were incubated with clofibrate (1 mM) for 6 h in the presence or absence of Baf. Pexophagy was assessed through the processing of RFP-GFP-SKL. (G) Control and PAN3 KD cells expressing pexophagy reporter SKL were treated with clofibrate (1 mM) for 6 h. GFP, RFP, and p62 signals were visualized and quantified by IF. Scale bars, 5 µM. (H) Control and KO HEK293A cells were incubated with either acute amino acid starvation (1.5 h, -AAa) or clofibrate (1 mM, 6 h). Whole-cell lysates were immunoblotted using the antibodies indicated. Unless otherwise indicated, experiments were performed three times. Data are represented as means ± SDs, and P values were determined by two-way ANOVA. *P ≤ 0.1; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001; ns, not significant. Source data are available for this figure: SourceData F5.

PAN3 activates pexophagy. (A) PAN3 is represented as a prominent teal dot on the volcano plot. Red dots on the graph denote common regulators across all four conditions. (B) Control and PAN3 KO HEK293A cells were treated with clofibrate (1 mM) for 6 h in the presence or absence of Baf. Changes in p62 and LC3B levels were analyzed using WB. (C) Control and KO cells were incubated with clofibrate (1 mM) for 6 h. p62 and LC3B puncta were visualized and quantified by IF. Scale bars, 10 µM. (D) Control and KO cells were treated with clofibrate in the presence or absence of Baf. WB was then used to examine the pexophagy receptor, PMP70. (E) Indicated cells were incubated with clofibrate. PMP70 signal was visualized and quantified by IF. Scale bars, 10 µM. (F) Control and PAN3 KD HCT116 cells stably expressing the RFP-GFP-SKL reporter were incubated with clofibrate (1 mM) for 6 h in the presence or absence of Baf. Pexophagy was assessed through the processing of RFP-GFP-SKL. (G) Control and PAN3 KD cells expressing pexophagy reporter SKL were treated with clofibrate (1 mM) for 6 h. GFP, RFP, and p62 signals were visualized and quantified by IF. Scale bars, 5 µM. (H) Control and KO HEK293A cells were incubated with either acute amino acid starvation (1.5 h, -AAa) or clofibrate (1 mM, 6 h). Whole-cell lysates were immunoblotted using the antibodies indicated. Unless otherwise indicated, experiments were performed three times. Data are represented as means ± SDs, and P values were determined by two-way ANOVA. *P ≤ 0.1; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001; ns, not significant. Source data are available for this figure: SourceData F5.

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