Journal: International journal of oncology

Article Title: A formulated red ginseng extract upregulates CHOP and increases TRAIL-mediated cytotoxicity in human hepatocellular carcinoma cells.

doi: 10.3892/ijo.2013.1964

Figure Lengend Snippet: Figure 4. The RGE sensitization to TRAIL-derived cell death occurs via death receptor 5 (DR5) upregulation in HCC cell lines. (A) DR5 protein levels were strongly dependent upon RGE concentration. (B) RGE‑induced upregulation of DR5, but not DR4, in HepG2 cells. (C) Upregulation of DR5 protein levels by RGE treatment in the different HCC cell lines.

Article Snippet: The following reagents were purchased and used according to the manufacturer's instructions: glutathione S-transferase (GST)-TRAIL and anti-DR5 antibodies were from Koma Biotechnologies (Seoul, Korea); anti-caspase 3, anti-PARP and anti-CHOP antibodies were from Cell Signaling Technology; anti-DR4 antibody was from Rockland; anti-tubulin antibody was from Abcam; anti-actin antibody, thapsigargin (Tg), necrostatin-1, NAC and BHA were from Sigma; and zVAD was from R&D Systems.

Techniques: Derivative Assay, Concentration Assay

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 TRAIL-death receptor signaling in VHL -deficient ccRCC cells (A) Volcano plot of SGI1027-induced and -repressed genes in RCC10 cells ( n = 2 biological replicates). FC, fold change. (B) Biocarta pathway enrichment analysis of SGI1027-induced genes in RCC10 cells. (C) RT-qPCR analysis of TNFSF10 , TNFRSF10A , TNFRSF10B , and TNFRSF10D mRNA levels in RCC10 cells treated with vehicle or SGI1027 for 2 days ( n = 3 biological replicates). (D and E) Immunoblot analysis of TRAIL, DR4, DR5, DcR2, pro-caspase-10, and cleaved caspase-10 (C-caspase-10) proteins in isogenic RCC10 cells treated with vehicle, SGI1027 (D and E, n = 2 biological replicates), MS1129 (E, n = 2 biological replicates), or decitabine (E, n = 2 biological replicates) for 2 or 7 days. (F) Global m5C levels in RCC10 cells treated with vehicle or SGI1027 for 2 days by ELISA assay ( n = 3 biological replicates). (G–J) MeDIP-qPCR assay in RCC10 cells treated with vehicle or SGI1027 for 2 days ( n = 3 biological replicates). (K–M) DNMT1, DNMT3A, and DNMT3B ChIP-qPCR assay in RCC10 cells ( n = 3 biological replicates). (N) Scheme of dDNMT-activated apoptotic pathway. Data represent mean ± SEM. p value was determined by bioinformatics with edgeR (A) or gene set enrichment analysis (B), unpaired 2-tailed Student’s t test (C and F), two-way ANOVA with Tukey’s test (G–I), and one-way ANOVA with Dunnett’s test (K–M). 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: Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Methylated DNA Immunoprecipitation, 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: Biochemistry and Biophysics Reports

Article Title: Core 2 β1,6-N-acetylglucosaminyltransferases accelerate the escape of choriocarcinoma from natural killer cell immunity

doi: 10.1016/j.bbrep.2021.100951

Figure Lengend Snippet: Association between C2GnT expression and NK cell toxicity via the TRAIL-DR pathways. (A) Viability of control, C2GnT KO Jar and BeWo cells using MTS assay. Data were obtained from three individual experiments and each condition was assessed by triplicates. Mean ± SD of relative cell counts are shown. (B) Cell viability of control, C2GnT KO Jar and BeWo cells treated with endo-β-galactosidase with 1 μg/mL TRAIL for 24 h. Data were obtained from three individual experiments and each condition was assessed by triplicates. Mean ± SD of cytotoxicity rates are shown. n.s., not significant. (C) Representative western blot demonstrating MUC1 and DR4 protein expression in control, C2GnT KO Jar and BeWo cells. LEL immunoprecipitates were subjected to western blotting with anti-MUC1.

Article Snippet: Western blotting was performed as previously described [ ] using the anti-C2GnT1 (Wako), anti-MICA/B (Biolegend), anti-death receptor 4 (DR4) (Funakoshi, Tokyo, Japan), and anti-mucin1 (MUC1) (Abcam, Cambridge, UK) antibodies.

Techniques: Expressing, Control, MTS Assay, Western Blot