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: dDNMT activates the apoptotic pathway in VHL -deficient ccRCC cells (A) Representative images of death of RCC10 cells pre-treated with vehicle, Z-VAD-FMK, necrostatin-1, ferrostatin-1, olaparib, 3-MA, or AC-FLTD-CMK for 30 min, followed by 3 days of co-treatment with SGI1027. (B) Quantification of PI-positive cells in (A) ( n = 3 biological replicates). (C) Representative images of death of RCC10 cells pre-treated with vehicle or Z-VAD-FMK for 30 min, followed by 3 days of co-treatment with MS1129. (D) Quantification of PI-positive cells in (C) ( n = 3 biological replicates). (E and F) Immunoblot analysis of C-caspase-3, C-caspase-7, and C-PARP1 proteins in RCC10 cells treated with vehicle, SGI1027 (E, n = 3 biological replicates), or MS1129 (F, n = 2 biological replicates) for 2 days. Data represent mean ± SEM. p value was determined by one-way ANOVA with Tukey’s test (B and D). Scale bars: 100 μm in (A and C). See also and .

Article Snippet: The following antibodies were used: anti-HIF-1α antibody (A300-286A, Bethyl Laboratories), anti-HIF-2α antibody (home-made), anti-DNMT1 antibody (24206-1-AP, Proteintech), anti-DNMT3A antibody (20954-1-AP, Proteintech), anti-DNMT3B antibody (26971-1-AP, Proteintech), anti-DNMT3L antibody (sc-393603, Santa Cruz), anti-cleaved-caspase-3 antibody (9661S, Cell Signaling Technology), anti-cleaved-caspase-7 antibody (8438S, Cell Signaling Technology), anti-PARP1 antibody (9542S, Cell Signaling Technology), anti-caspase-10 antibody (14311-1-AP, Proteintech), anti-caspase-8 antibody (13423-1-AP, Proteintech), anti-TRAIL antibody (27064-1-AP, Proteintech), anti-DR4 antibody (24063-1-AP, Proteintech), anti-DR5 antibody (69400S, Cell Signaling Technology), anti-DcR2 antibody (16781-1-AP, Proteintech), anti-β-actin (81115-1-RR, Proteintech), and anti-α-tubulin (66031-1-Ig, Proteintech).

Techniques: 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: The following antibodies were used: anti-HIF-1α antibody (A300-286A, Bethyl Laboratories), anti-HIF-2α antibody (home-made), anti-DNMT1 antibody (24206-1-AP, Proteintech), anti-DNMT3A antibody (20954-1-AP, Proteintech), anti-DNMT3B antibody (26971-1-AP, Proteintech), anti-DNMT3L antibody (sc-393603, Santa Cruz), anti-cleaved-caspase-3 antibody (9661S, Cell Signaling Technology), anti-cleaved-caspase-7 antibody (8438S, Cell Signaling Technology), anti-PARP1 antibody (9542S, Cell Signaling Technology), anti-caspase-10 antibody (14311-1-AP, Proteintech), anti-caspase-8 antibody (13423-1-AP, Proteintech), anti-TRAIL antibody (27064-1-AP, Proteintech), anti-DR4 antibody (24063-1-AP, Proteintech), anti-DR5 antibody (69400S, Cell Signaling Technology), anti-DcR2 antibody (16781-1-AP, Proteintech), anti-β-actin (81115-1-RR, Proteintech), and anti-α-tubulin (66031-1-Ig, Proteintech).

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

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: dDNMT specifically kills patient-derived VHL -deficient ccRCC in mice (A) Tumor growth curves of VHL -deficient UTSW-PDX206, UTSW-PDX258, UTSW-PDX490, and UTSW-PDX26 in mice treated with vehicle (Veh) or SGI1027 for 10 days. (B) Tumor growth curves of VHL -WT UTSW-PDX416 and UTSW-PDX143 in mice treated with vehicle or SGI1027 for 10 days. (C) Kaplan-Meier survival curve of UTSW-PDX490-bearing mice ( n = 10 biological replicates). (D and E) Global m5C levels in UTSW-PDX206 (D) or UTSW-PDX258 (E) tumors harvested from mice after treatments by ELISA assay ( n = 5 biological replicates). (F) Immunoblot analysis of DNMT1, DNMT3A, DNMT3B, TRAIL, DR4, DR5, procaspase-10, C-caspase-3, and C-caspase-7 proteins in UTSW-PDX258 tumors harvested from mice after treatments ( n = 5 biological replicates). (G) Representative C-caspase-3 IHC in UTSW-PDX258 tumors. Scale bar, 100 μm. (H) Quantification of C-caspase-3-positive cells in (G) ( n = 5 biological replicates). (I) Immunoblot analysis of DNMT1, DNMT3A, DNMT3B, TRAIL, DR4, DR5, C-caspase-3, and VHL proteins in UTSW-PDX416 tumors harvested from mice after treatments ( n = 5 biological replicates). (J) Immunoblot analysis of DNMT1, DNMT3A, DNMT3B, and procaspase-10 proteins in UTSW-PDX206, UTSW-PDX258, and UTSW-PDX26 tumors ( n = 4–5 biological replicates). Data represent mean ± SEM. p value was determined by two-way ANOVA with Tukey’s test (A), log rank test (C), and unpaired 2-tailed Student’s t test (D, E, and H). See also and ; .

Article Snippet: The following antibodies were used: anti-HIF-1α antibody (A300-286A, Bethyl Laboratories), anti-HIF-2α antibody (home-made), anti-DNMT1 antibody (24206-1-AP, Proteintech), anti-DNMT3A antibody (20954-1-AP, Proteintech), anti-DNMT3B antibody (26971-1-AP, Proteintech), anti-DNMT3L antibody (sc-393603, Santa Cruz), anti-cleaved-caspase-3 antibody (9661S, Cell Signaling Technology), anti-cleaved-caspase-7 antibody (8438S, Cell Signaling Technology), anti-PARP1 antibody (9542S, Cell Signaling Technology), anti-caspase-10 antibody (14311-1-AP, Proteintech), anti-caspase-8 antibody (13423-1-AP, Proteintech), anti-TRAIL antibody (27064-1-AP, Proteintech), anti-DR4 antibody (24063-1-AP, Proteintech), anti-DR5 antibody (69400S, Cell Signaling Technology), anti-DcR2 antibody (16781-1-AP, Proteintech), anti-β-actin (81115-1-RR, Proteintech), and anti-α-tubulin (66031-1-Ig, Proteintech).

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Clinical and Translational Medicine

Article Title: NLK facilitates Caspase‐8 activation to drive macrophage PANoptosis in sepsis

doi: 10.1002/ctm2.70616

Figure Lengend Snippet: LPS stimulation induces PANoptosis in BMDMs. (A) Experimental scheme for BMDMs generation and LPS stimulation. (B, C) Representative time‐course immunoblot analysis of NLK and PANoptosis‐related markers, including apoptosis (cleaved Caspase‐3, Caspase‐7), necroptosis (MLKL, p‐MLKL), and pyroptosis (NLRP3, pro‐/cleaved Caspase‐1, full‐length/N‐terminal GSDMD) in WT BMDMs following LPS treatment. (D, E) PI/calcein AM staining of BMDMs at the indicated time points (calcein AM for viable cells, green; PI for dead cells, red; scale bar: 100 µm) and quantitative analysis ( n = 3 per group, independent biological replicates). (F) Time‐course immunoblot analysis of NLK, Caspase‐8, cleaved Caspase‐3, and full‐length/N‐terminal GSDMD in WT and NKO BMDMs following LPS treatment. (G–K) Quantitative analysis of NLK, Caspase‐8, cleaved Caspase‐3, and full‐length/N‐terminal GSDMD in panel F ( n = 3 independent biological replicates). Statistical significance was determined using one‑way ANOVA with Bonferroni's post hoc test; * p < .05 and ** p < .01.

Article Snippet: Primary antibodies against NLK (#94350), Caspase‐8 (#8592), Caspase‐3 (#96662), Caspase‐7 (#9492), MLKL (#37705), p‐MLKL (#37333), RIPK3 (#95702), p‐RIPK3 (#91702S), RIPK1 (#3493), NLRP3 (#15101), GSDMD (#39754), and ASC (#67824) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Western Blot, Staining

Journal: Clinical and Translational Medicine

Article Title: NLK facilitates Caspase‐8 activation to drive macrophage PANoptosis in sepsis

doi: 10.1002/ctm2.70616

Figure Lengend Snippet: NLK deficiency shifts LPS‐induced PANoptosis towards necroptosis in BMDMs. (A, B) Representative immunoblots of pyroptosis (NLRP3, Caspase‐1, GSDMD), apoptosis (Caspase‐3, Caspase‐7), and necroptosis (MLKL, p‐MLKL) markers in PBS/LPS‐treated BMDMs. (C, D) Immunofluorescent co‐staining and quantification of CD68 (green) with GSDMD‐N (purple, pyroptosis), cleaved caspase‐3 (orange, apoptosis), and p‐MLKL (red, necroptosis) in BMDMs from each group. Scale bar: 100 µm; n = 3 independent biological replicates. (E) Representative TEM images of BMDMs showing the ultrastructural features consistent with pyroptosis, apoptosis, and necroptosis. Blue arrows indicate chromatin condensation and nuclear shrinkage (apoptosis); red arrows and circles denote cellular swelling, membrane pore formation and vesiculation (pyroptosis); yellow arrows indicate plasma membrane rupture and translucent cytoplasm (necroptosis). Scale bar: 0.1 µm. (F–L) Quantification of immunoblot signals from panels A and B. (M) Relative proportions of BMDMs exhibiting predominant apoptosis‐, pyroptosis‐, or necroptosis‐associated ultrastructural features based on TEM. Each bar represents the distribution of ultrastructural phenotypes among 100 macrophages analysed per sample from n = 3 independent biological replicates; cells lacking overt death‐associated features or displaying ambiguous morphology were classified as unclassified. Statistical differences were analysed by one‑way ANOVA with Bonferroni's post hoc test, * p < .05 and ** p < .01.

Article Snippet: Primary antibodies against NLK (#94350), Caspase‐8 (#8592), Caspase‐3 (#96662), Caspase‐7 (#9492), MLKL (#37705), p‐MLKL (#37333), RIPK3 (#95702), p‐RIPK3 (#91702S), RIPK1 (#3493), NLRP3 (#15101), GSDMD (#39754), and ASC (#67824) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Western Blot, Staining, Membrane, Clinical Proteomics

Journal: Cancer Cell International

Article Title: Targeted suppression of DNA polymerase θ enhances calcium flux and triggers intrinsic apoptosis in gastric cancer

doi: 10.1186/s12935-026-04206-5

Figure Lengend Snippet: NVB S suppresses HGC-27 xenograft tumor growth without causing systemic toxicity. (A) Schematic illustration of the experimental timeline and treatment regimen. (B) Representative photographs of excised tumors from each treatment group at the study endpoint (day 14; n = 7 mouse per group). (C) Tumor growth curves showing volume measurements over time for each group. (D) Quantitative analysis of final tumor weights across treatment groups. (E) Representative immunohistochemical images of tumor sections showing NVB S treatment inhibits tumor progression in mice, with indicators including: (i) H&E staining, (ii) Ki67, (iii) cleaved caspase-3/7, (iv) γ-H2AX and TUNEL, (v) RAD51 staining (scale bar, 50 μm, n = 5 independent experiments). (F) Western blot analysis of γ-H2AX and cleaved caspase-7 in tumor lysates from each group ( n = 3 independent experiments). (G) Western blot analysis of p-RPA32 (Ser8) and total RPA32 expression in tumor lysates from each group ( n = 3 independent experiments). (H) Western blot analysis of p-CaMKII/CaMKII expression in tumor lysates from each group ( n = 3 independent experiments). (Low: NVB S 50 mg/kg; High: 100 mg/kg). All data are presented as mean ± SD. Statistical significance was assessed using One-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001, N.S., not significant

Article Snippet: The membranes were blocked with 5% non-fat milk (Yili, Hohhot, China) in TBST (Tris-buffered saline with Tween-20) for 1 h at room temperature and incubated overnight at 4 °C with primary antibodies against POLQ (1:1000, Cat. No. SAB1402530, Sigma-Aldrich, Germany), β-actin (1:10,000, Cat. No. EM21002, Hua’an Biotechnology Co., Ltd., Hangzhou, China), cleaved caspase-3 (1:500, Cat. No. 9661 S, Cell Signaling Technology, USA), cleaved caspase-7 (1:500, Cat. No. 8438 S, Cell Signaling Technology, USA),γ-H2AX (1:1000, Cat. No. AB81299, Abcam, UK), RPA32 (1:10,000, Cat. No. 67999-1-Ig, proteintech, China).

Techniques: Immunohistochemical staining, Staining, TUNEL Assay, Western Blot, Expressing