Research Paper Volume 15, Issue 19 pp 10407—10427

Figure 6. PLCL1 promotes autophagy by interacting with DEPP. (A) Volcano plot of autophagy-related DEGs (eight candidates, black colour) in 786-O cells with PLCL1 overexpression and vector cell sequencing. The red dots in the volcano represent upregulated DEGs, and the green dots represent downregulated DEGs, while black dots represent non-significant DEGs. (B) The association between PLCL1 and DEPP was calculated using Pearson analysis. (C, D) Wound healing assays were used to determine the role of DEPP in RCC in vitro and quantification analyses of the results. Scale bar: 100 μm. (E) Transwell assays were performed in 786-O and ACHN cells transfected with DEPP or vector control. Scale bar: 100 μm. (F–H) Western blots of LC3B and P62 were performed in 786-O and ACHN cells transfected with DEPP or vector control. (I) 786-O and ACHN cells were immunofluorescently stained with PLCL1 (green) and DEPP (red) antibodies to assess colocalization using a confocal microscope. Scale bar, 20 μm. (J, K) Interaction between PLCL and DEPP in 786-O cells. The coimmunoprecipitates were utilized for western blotting with anti-PLCL1 and anti-MYC antibodies. Data are shown as the mean ± SE from three independent experiments. Student’s t test was performed to determine statistical significance between two groups. ^*P < 0.05; ^**P < 0.01; ^***P < 0.001 versus vector group.