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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 <t>bioactive</t> 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 .
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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 <t>bioactive</t> 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 .
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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 <t>bioactive</t> 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 .
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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 <t>bioactive</t> 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 .
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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 <t>bioactive</t> 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 .
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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 <t>bioactive</t> 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 .
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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 .

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: Bioactive Compound Library , Selleckchem , Cat# L1700.

Techniques: In Vitro, Western Blot