Figure 3 from Oncogenic Mutant p53 Sensitizes Non–Small Cell Lung Cancer Cells to Proteasome Inhibition via Oxidative Stress–Dependent Induction of Mitochondrial Apoptosis
Year: 2024
Type: other
Abstract: BTZ induces ATF3/NOXA and caspase-dependent apoptosis. A, H1975 cells were pretreated (30 minutes) with vehicle or the pan-caspase inhibitor QVD-OPh (QVD; 1 µmol/L) and then treated with vehicle or BTZ (5 nmol/L) for 48 hours. Cell viability was determined by trypan blue exclusion assay. B, H1975 cells were treated as in A, and the percentage of apoptotic cells was determined by annexin V–propidium iodide staining followed by FACS analysis. C, Cell lysates from A were immunoblotted with the indicated antibodies. D, H1975 or H460 cells were treated with vehicle or 2 or 5 nmol/L of BTZ for 48 hours, and cell lysates were immunoblotted with the indicated antibodies. E, RNA from D was extracted and subjected to qRT-PCR with NOXA primers. F, H1975 cells stably expressing shC or shNOXA were treated with vehicle or BTZ (5 nmol/L) for 48 hours, and cell lysates were immunoblotted with the indicated antibodies. G, H1975 cells were treated with vehicle or 2 or 5 nmol/L of BTZ for 48 hours, and mRNA expression of ATF3 was analyzed by qRT-PCR. H, H1975 cells were treated with vehicle, NAC (1 mmol/L), BTZ (5 nmol/L), or BTZ + NAC for 48 hours, and cell lysates were immunoblotted with indicated antibodies. I, H1975 cells stably expressing control shRNA (shC) or ATF3 shRNA (shATF3) were treated with vehicle or BTZ (5 nmol/L) for 48 hours, and cell lysates were immunoblotted with the indicated antibodies. J, H1975 cells stably expressing shC or shATF3 were treated with vehicle or BTZ (5 nmol/L) for 48 hours, and the percentage of apoptotic cells was determined by annexin V–propidium iodide staining followed by FACS analysis. **, P < 0.01; ***, P < 0.005; ns indicates P > 0.05. Error bars indicate ± 1.0 SD. Cl. Casp 3, cleaved caspase-3.
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
OpenAlex ID: W4403432900