Journal: bioRxiv

Article Title: Activation of NLRP3 inflammasome/pyroptosis by protease inhibitor atazanavir at high concentrations is mediated through mitochondrial dysfunction

doi: 10.1101/2024.06.18.599585

Figure Lengend Snippet: A. Effect of ATV on cell viability. CEM cells were cultured with dimethyl sulfoxide (DMSO) and different concentrations of ATV (2.5, 5.0, 7.5, 15, 22.5 and 30.0 µM). At days 2, 4, and 8, aliquots of the cell culture were collected, and live cells were counted with trypan blue staining using hemocytometer. B. Apoptosis was assessed by staining with an FITC Annexin V Apoptosis Detection Kit I. Bar graphs represent proportion of apoptotic cells after 2, 4, and 8 days of treatment compared to DMSO-treated cells. C. Heatmap of RT 2 Profiler PCR Arrays result. Cells collected under DMSO and 30 µM ATV treatment at day 8 were used for comparison. D. mRNA expression levels of results of caspase-1 (CASP1), BCL2A1, tumor protein p73 (TP73) and TNFRSF1B by qPCR. Primers used are listed in . Data represent at least 3 independent experiments and plotted as mean ± SD. P-values are two sided and considered significant if <0.05 (*), < 0.01 (**), or <0.001 (***).

Article Snippet: To identify genes of interest, a 96 well RT 2 profiler PCR array human apoptosis kit (Qiagen, Germantown, MD) containing 84 apoptotic genes was used according to manufacturer’s instructions.

Techniques: Cell Culture, Staining, Comparison, Expressing

Journal: bioRxiv

Article Title: Activation of NLRP3 inflammasome/pyroptosis by protease inhibitor atazanavir at high concentrations is mediated through mitochondrial dysfunction

doi: 10.1101/2024.06.18.599585

Figure Lengend Snippet: CEM cells were cultured with dimethyl sulfoxide (DMSO) and different concentrations of ATV (2.5, 5.0, 7.5, 15, 22.5 and 30.0 µM) for 8 days. At day 8, the cells were harvested, and RNA and protein were extracted. CEM cells were pre-incubated with AC-YVAD-AMK (20 µM) or MCC950 (1 µM) for 1 h before ATV treatment. A. mRNA expression level of key genes in NLRP3 inflammasome pathway. Real time qPCR of NLRP3, ASC, IL-1β, IL-6, IL-18 were performed. The expression levels of the genes were normalized to the expression of housekeeping gene, GAPDH. Data (mean ± SD, n = 3), are represented as fold change in expression compared with cells treated with DMSO. B. Cells treated for 6 h were harvested, and damage or dead cells was assessed by staining with an FITC Annexin V Apoptosis Detection Kit I. Bar graphs represent proportion of damage or dead cells of ATV treatment compared to DMSO-treated cells. C. Western blot result of pro-caspase-1. The density of the pro-caspase-1 was normalized to that of GAPDH. D-H. cells after treating 6 h were harvested and RNA were extracted. mRNA expression level of NLRP3 (D), ASC (E), GSDMD (F), Caspase-1 (F) and IL-1β (H) by RT-qPCR. Data (mean ±SD) were compared with DMSO treated cells and analyzed by Student’s t-test. P-values are two sided and considered significant if <0.05 (*), < 0.01 (**), or <0.001 (***).

Article Snippet: To identify genes of interest, a 96 well RT 2 profiler PCR array human apoptosis kit (Qiagen, Germantown, MD) containing 84 apoptotic genes was used according to manufacturer’s instructions.

Techniques: Cell Culture, Incubation, Expressing, Staining, Western Blot, Quantitative RT-PCR

Journal: Current Issues in Molecular Biology

Article Title: DNA Double-Strand Break Repair Inhibitors: YU238259, A12B4C3 and DDRI-18 Overcome the Cisplatin Resistance in Human Ovarian Cancer Cells, but Not under Hypoxia Conditions

doi: 10.3390/cimb45100500

Figure Lengend Snippet: Genes analyzed with the RT 2 Profiler PCR Array—Human DNA Damage Signaling Pathway kit (Qiagen, Germantown, MD, USA).

Article Snippet: Expression of genes related to cell response to DNA damage was carried out using Real-time PCR using the RT 2 Profiler PCR Array—Human DNA Damage Signaling Pathway gene expression evaluation kit (Qiagen, Germantown, MD, USA).

Techniques: