Fig. 3: LDAH knockdown inhibits ER stress-mediated apoptosis in ovarian cancer cells. | Oncogenesis

Fig. 3: LDAH knockdown inhibits ER stress-mediated apoptosis in ovarian cancer cells.

From: Lipid droplet-associated hydrolase (LDAH) knockdown enhances TAG hydrolysis and promotes ovarian cancer progression and chemoresistance

Fig. 3

A Immunofluorescence images of DsRed2-ER5 (red) in LDAH-GFP expressing ES2 cells. Blue, DAPI. Scale bars, 20 µm. B Western blot of ER stress markers (Ero 1a, IREI, PDI, PERK) in scrambled control and LDAH knockdown ES2 cells, with or without cisplatin treatment. The quantification of band intensity normalized to GAPDH is shown on the right. Two-way ANOVA followed by Tukey’s multiple comparison test. C Immunofluorescence images of PDI (red) in scrambled control and LDAH knockdown ES2, with or without cisplatin treatment. Blue, DAPI. Scale bars, 50 µm. D Western blot of Bak1 in scrambled control and LDAH knockdown ES2cells, with or without cisplatin treatment. The quantification of band intensity normalized to GAPDH is shown on the right. One-way ANOVA followed by Tukey’s multiple comparison test. E, F qPCR analysis showing mRNA expression of Bak1 (E; pro-apoptotic) and BIRC2 (F; anti-apoptotic) in scrambled control and LDAH knockdown ES2, with and without cisplatin treatment. One-way ANOVA followed by Tukey’s multiple comparison test. G Immunofluorescence images of LDAH and PERK in scrambled control and LDAH overexpressing ES2 cancer cells. Scale bars, 20 µm. Quantification of PERK signal is shown on the right. Student’s T-test. H Western blot of cleaved caspase 3, p-PERK, PERK, and LDAH in scrambled and LDAH overexpressing cells after treatment with vehicle, cisplatin alone, or in combination with a PERK inhibitor GSK2606414. GAPDH, a loading control. The quantification of band intensity normalized to GAPDH is shown on the right. One-way ANOVA followed by Tukey’s multiple comparison test. I mRNA expression level of Bak1 (pro-apoptosis) in LDAH overexpressing cells after treatment with vehicle, cisplatin alone or combination of in combination with a PERK inhibitor GSK2606414. One-way ANOVA followed by Tukey’s multiple comparison test. Error bars, SD. *, p < 0.05, **, p < 0.01, ***, p < 0.001. ****, p < 0.0001. ns, not significant.

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