Journal: Cell Reports Medicine

Article Title: HIF-activated priming of TRAIL-induced cell death determines epigenetic vulnerability in kidney cancer

doi: 10.1016/j.xcrm.2026.102630

Figure Lengend Snippet: In vitro chemical screens identify SGI1027 and MS1129 specifically killing VHL -deficient ccRCC cells in an HIF-dependent manner (A) Scheme of cell viability screens with 2,645 FDA-approved drugs and bioactive compounds in isogenic ccRCC cells. (B) The cell viability ratio of all chemicals (5 μM) was ranked from the primary screen ( n = 1 biological replicate). (C) The IC50 ratio of 23 positive hits in the secondary screen ( n = 1 biological replicate). (D) Representative images of death of parental and HIF-1α/2α-DKO RCC10 cells treated with vehicle or SGI1027 for 3 days. BF, bright field. (E) Quantification of PI-positive cells in (D) ( n = 3 biological replicates). (F) Immunoblot analysis of HIF-1α and HIF-2α proteins in isogenic RCC10 cells ( n = 3 biological replicates). (G) The IC50 of SGI1027 in isogenic RCC10 and HIF-1α- and/or HIF-2α-ΚΟ cells ( n = 3 biological replicates). (H) Clonogenic growth of isogenic RCC10 and HIF-1α- and/or HIF-2α-ΚΟ cells treated with vehicle or SGI1027 for 9 days. (I) Quantification of crystal violet intensity in (H) ( n = 3 biological replicates). (J) The cell viability ratio of SGI1027 analogs (2 μM) in parental and HIF-1α/2α-DKO RCC10 cells ( n = 1 biological replicate). (K) Representative images of death of parental and HIF-1α/2α-DKO RCC10 cells treated with vehicle or MS1129 for 3 days. (L) Quantification of PI-positive cells in (K) ( n = 3 biological replicates). (M) The IC50s of SGI1027, MS1129, and MS1143 in isogenic RCC10 and RCC10-HIF-1α/2α-DKO cells ( n = 3 biological replicates). dDNMT, DNMT protein degrader. Data represent mean ± SEM. p value was determined by two-way ANOVA with Tukey’s test (E, I, and L). Scale bars, 100 μm in (D) and (K). See also and ; and .

Article Snippet: anti-HIF-1α antibody , Bethyl Laboratories or home-made , A300-286A, RRID: AB_2117114.

Techniques: In Vitro, Western Blot

Journal: Cell Reports Medicine

Article Title: HIF-activated priming of TRAIL-induced cell death determines epigenetic vulnerability in kidney cancer

doi: 10.1016/j.xcrm.2026.102630

Figure Lengend Snippet: HIF-induced procaspase-10 predisposes VHL -deficient ccRCC to apoptosis upon dDNMT treatment (A) RT-qPCR analysis of CASP10 mRNA levels in isogenic RCC10 cells ( n = 3 biological replicates). (B) CASP10 mRNA levels in human VHL -WT and mutant ccRCC. Data were retrieved from TCGA, firehose Legacy in cBioPortal. (C) Immunoblot analysis of HIF-1α, HIF-2α, and procaspase-10 proteins in isogenic RCC10 cells. (D) Nucleotide sequence of the hypoxia response element ([HRE], in blue) at the promoter of the CASP10 gene (top). Genome browser snapshot of HIF-1α ChIP-seq peaks detected in hypoxic MDA-MB-231 cells (blue, n = 2 biological replicates), and HIF-2α ChIP-seq peaks detected 786-O cells (red, n = 2 biological replicates, GSE253325 ). (E) ChIP-qPCR assay showing enrichment of HIF-1α and HIF-2α at the promoter of CASP10 in RCC10 cells ( n = 3 biological replicates). (F) Representative images of death of RCC10 cells pre-treated with vehicle or CASP10i Z-AEVD-FMK for 30 min, followed by 3 days of co-treatment with SGI1027. (G) Quantification of PI-positive cells in (F) ( n = 3 biological replicates). (H) Representative images of death of RCC10 cells pre-treated with vehicle or CASP10i Z-AEVD-FMK for 30 min, followed by 3 days of co-treatment with MS1129. (I) Quantification of PI-positive cells in (H) ( n = 3 biological replicates). (J and K) Immunoblot analysis of procaspase-10, C-caspase-3, and C-caspase-7 proteins in RCC10 cells pre-treated with vehicle or CASP10i Z-AEVD-FMK for 30 min, followed by 2 days of co-treatment with SGI1027 (J, n = 2 biological replicates) or MS1129 (K, n = 2 biological replicates). Data represent mean ± SEM. p value was determined by one-way ANOVA with Dunnett’s test (A and E), unpaired 2-tailed Student’s t test (B), and one-way ANOVA with Tukey’s test (G and I). Scale bars: 100 μm in (F and H). See also .

Article Snippet: anti-HIF-1α antibody , Bethyl Laboratories or home-made , A300-286A, RRID: AB_2117114.

Techniques: Quantitative RT-PCR, Mutagenesis, Western Blot, Sequencing, ChIP-sequencing, ChIP-qPCR