Journal: Apoptosis : an international journal on programmed cell death

Article Title: Targeting PNPO to suppress tumor growth via inhibiting autophagic flux and to reverse paclitaxel resistance in ovarian cancer.

doi: 10.1007/s10495-024-01956-3

Figure Lengend Snippet: Fig. 3 Effect of PNPO on the biogenesis and distribution of lysosomes in ovarian cancer cells. A Detection of lysosomes in OVCAR-3 and SK-OV-3 cells. Cells were plated into a 6-well plate and transiently transfected with PNPO-overexpressing plasmid for 48 h. The lysosomes were detected by the LysoTracker assay using red acidic probes. Scale bar, 25 μm. B Detection of lysosome-related mRNAs by qRT-PCR in cells after PNPO-overexpressing plasmid transfection. C Detection of lysosome-related mRNAs by qRT-PCR in cells after PNPO si- RNA transfection. D Correlation of PNPO with LAMP2 using the GEPIA2 (http://gepia.cancer-pku. cn). PNPO expression was posi tively correlated with LAMP2 expression. E Effect of PNPO on LAMP2 mRNA expression in ovarian cancer cells after PNPO-overexpressing plasmid transfection. F Effect of PNPO on LAMP2 mRNA expression in ovarian cancer cells after PNPO si-RNA transfection. G Detection of LAMP2 protein by immuno fluorescence staining. PNPO- overexpressing cells were plated into a confocal dish for 48 h and LAMP2 was detected using the immunofluorescent assay. Scale bar, 100 μm. All assays were repeated at least three times. Data were presented as mean ± SD. P values were calculated by the two-sided unpaired Student’s t-test. oe-PNPO, PNPO-over expressing plasmid; si-PNPO, PNPO si-RNA; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, p < 0.0001

Article Snippet: After blocking with 10% normal goat serum for 40 min at room temperature, the section was incubated with an anti-PNPO antibody (1:400 dilution, Proteintech), anti-Ki67 antibody (Proteintech), anti-LAMP2 antibody (1:600 dilution, Proteintech), 1 3

Techniques: Transfection, Plasmid Preparation, Quantitative RT-PCR, Expressing, Fluorescence, Staining

Journal: Apoptosis : an international journal on programmed cell death

Article Title: Targeting PNPO to suppress tumor growth via inhibiting autophagic flux and to reverse paclitaxel resistance in ovarian cancer.

doi: 10.1007/s10495-024-01956-3

Figure Lengend Snippet: Fig. 5 Effect of LAMP2-siRNA in OVCAR-3 and SK-OV-3 cells. A Detection of LAMP2 mRNA by qRT-PCR. Knockdown of LAMP2 was validated in cells transfected with 3 LAMP2- siRNAs for 48 h. B Detection of LAMP2 protein by Western blot in cells transfected with 2 LAMP2-siRNAs for 48 h. C Measurement of cell viability by the CCK8 assay. Cells were seeded into a 96-well plate and were transfected with negative control-siRNA or si-LAMP2 for 24 h. Silencing LAMP2 suppressed OC cell growth. D-G Detection of apoptosis. Cells were transfected with LAMP2- siRNA for 48 h and apoptotic cells were measured using flow cytometry. Data were presented as mean ± SD (n = 3). P values were calculated by the one-way ANOVA followed by Tukey’s multiple comparisons test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001

Article Snippet: After blocking with 10% normal goat serum for 40 min at room temperature, the section was incubated with an anti-PNPO antibody (1:400 dilution, Proteintech), anti-Ki67 antibody (Proteintech), anti-LAMP2 antibody (1:600 dilution, Proteintech), 1 3

Techniques: Quantitative RT-PCR, Knockdown, Transfection, Western Blot, CCK-8 Assay, Negative Control, Flow Cytometry