Journal: International Journal of Molecular Sciences

Article Title: 6-Shogaol Overcomes Gefitinib Resistance via ER Stress in Ovarian Cancer Cells

doi: 10.3390/ijms24032639

Figure Lengend Snippet: 6-shogaol induces the binding of CHOP on DR5 promoter in ovarian cancer cells. ( A – D ) After A2780 and OVCAR-3 cells were transfected with CHOP siRNA (30 nM, 24 h), WST-1, intracellular Ca 2+ assay, Western blotting analyses, and ChIP assay were performed with/without 6-shogaol (30 μM, 24 h) treatment.; *, p < 0.05. Western blotting analyses were carried out to determine the expression of CHOP, DR4, DR5, and caspase-3 cleavage in 6-shogaol-treated CHOP knockdown cells. β-actin was used as a protein loading control. ( E – H ) After A2780 and OVCAR-3 cells were transfected with DR5 (30 nM, 24 h), WST-1, LDH cytotoxicity assay, intracellular Ca 2+ assay, and Western blotting analyses were performed with/without 6-shogaol (30 μM, 24 h) treatment.; *, p < 0.05. n.s = not significant. Western blotting was carried out to identify the expression of DR5, caspase-3, and caspase-8 cleavage in 6-shogaol-treated DR5 knockdown cells. β-actin was used as a protein loading control.

Article Snippet: The primary antibodies used included β-actin (Santa Cruz, 1:1000, sc-47778), eIF2α (Santa Cruz, 1:1000, sc-133132), GRP78 (Santa Cruz, 1:1000, sc-166490); CD63 (Abcam, 1:1000, ab216130); Nox4 (Proteintech, 1:1000, 14347-1-AP); and cleaved caspase-3 (Cell Signaling, 1:1000, #9664), cleaved caspase-8 (Cell Signaling, 1:1000, #9748), cleaved caspase-9 (Cell Signaling, 1:1000, #20750), p-PERK(Thr980) (Cell Signaling, 1:1000, #3179), PERK (Cell Signaling, 1:1000, #5683), p-eIF2α (Ser51) (Cell Signaling, 1:1000, #3398), ATF4 (Cell Signaling, 1:1000, #11815), CHOP (Cell Signaling, 1:1000, #2895), DR4 (Cell Signaling, 1:1000, #42533), DR5 (Cell Signaling, 1:1000, #8074), E-cadherin (Cell Signaling, 1:1000, #14472), N-cadherin (Cell Signaling, 1:1000, #13116), Slug (Cell Signaling, 1:1000, #9585), Snail (Cell Signaling, 1:1000, #3879), and vimentin (CellSignaling, 1:1000, #5741).

Techniques: Binding Assay, Transfection, Western Blot, Expressing, Knockdown, Control, LDH Cytotoxicity Assay

Journal: Molecular Cell

Article Title: The TRAIL-Induced Cancer Secretome Promotes a Tumor-Supportive Immune Microenvironment via CCR2

doi: 10.1016/j.molcel.2017.01.021

Figure Lengend Snippet:

Article Snippet: α-TRAIL-R1 , ProSci , Cat#PSC-1139-C100.

Techniques: Western Blot, Flow Cytometry, Recombinant, In Vivo, Red Blood Cell Lysis, Blocking Assay, Enzyme-linked Immunosorbent Assay, Proliferation Assay, cDNA Synthesis, Sequencing, Plasmid Preparation, Control, shRNA, Software

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: BMC Cancer

Article Title: Snake venom toxin from vipera lebetina turanica induces apoptosis of colon cancer cells via upregulation of ROS- and JNK-mediated death receptor expression

doi: 10.1186/1471-2407-12-228

Figure Lengend Snippet: Effect of snake venom toxin on ROS generation and the expression of death receptors in human colon cancer cells. a, Effect of snake venom toxin on ROS generation by treatment of snake venom toxin in colon cancer cells. After treatment of snake venom toxin for 30 min, the cells were incubated with 10 μM DCF-DA at 37°C for 4 h, and then washed twice with PBS. The fluorescence intensity of DCF was measured in a microplate-reader at an excitation wavelength of 485 nm and an emission wavelength of 538 nm. b , Two colon cancer cells, HCT116 cells and HT-29 cells were treated with snake venom toxin (0.1, 0.5, 1 μg/ml) at 37°C for 24 h, and equal amounts of total proteins (50 μg/lane) were subjected to 12% SDS-PAGE. Expression of DR4, DR5 and β-actin was detected by Western blotting using specific antibodies. β-actin protein was used an internal control. Each band is representative for three experiments. Columns, means of three experiments, with triplicates of each experiment; bars, SD. *, p <0.05, significantly different from non treated control group.

Article Snippet: Small interfering (si) RNA species for death receptor (DR4 and DR5) and non-targeting control siRNA were purchased from Bioneer (Daejeon, Korea), and death receptor 4 (DR4) was purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA)

Techniques: Expressing, Incubation, Fluorescence, SDS Page, Western Blot, Control

Journal: BMC Cancer

Article Title: Snake venom toxin from vipera lebetina turanica induces apoptosis of colon cancer cells via upregulation of ROS- and JNK-mediated death receptor expression

doi: 10.1186/1471-2407-12-228

Figure Lengend Snippet: Effects of DR4 or DR5 knockdown on snake venom toxin induced cell viability inhibition and caspase-3 activation. a, HCT116 cells and HT-29 cells were transfected with non targeting control siRNA or DR4 or DR5 siRNA (100 nM) as described in Methods for 24 h. Then, implemented snake venom toxin was treated (1 μg/ml) for another 24 h. Thereafter, cell viability was measured by direct counting after trypan blue staining. b, Equal amounts of total proteins (50 μg/lane) were subjected to 12% SDS-PAGE. Expression of DR4, DR5, cleaved caspase-3 and β-actin was detected by Western blotting using specific antibodies. β-actin protein was used an internal control. Each band is representative for three experiments. Columns, means of three experiments, with triplicates of each experiment; bars, SD. *, p <0.05, significantly different from non treated control group. #, p <0.01 significantly different from sc siRNA -treated group.

Article Snippet: Small interfering (si) RNA species for death receptor (DR4 and DR5) and non-targeting control siRNA were purchased from Bioneer (Daejeon, Korea), and death receptor 4 (DR4) was purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA)

Techniques: Knockdown, Inhibition, Activation Assay, Transfection, Control, Staining, SDS Page, Expressing, Western Blot

Journal: BMC Cancer

Article Title: Snake venom toxin from vipera lebetina turanica induces apoptosis of colon cancer cells via upregulation of ROS- and JNK-mediated death receptor expression

doi: 10.1186/1471-2407-12-228

Figure Lengend Snippet: Effect of JNK pathway on the upregulation of DR4 or DR5, and cell death by snake venom toxin. a , Effect of snake venom toxin on the expression of MAPK proteins in colon cancer cells. HCT116 cells and HT-29 cells were treated with snake venom toxin for 24 h and whole cell extracts were analyzed by western blotting using the relevant antibodies. b , Effect of SP600125 on the cell viability in snake venom toxin treated cancer cells. Cells were pretreated with SP600125 (0, 5, 10 μM) for 1 h and then treated with snake venom toxin for 24 h. The results were expressed as a percentage of viable cells. c , Effect of JNK inhibitor (SP600125) on the expression of death receptors. Cells were pretreated with SP600125 (10 μM) for 1 h, and then cells were treated with snake venom toxin for 24 h, and whole cell extracts were analyzed by Western blotting using DR4, DR5, p-JNK and β-actin antibodies. Each band is representative for three experiments. Columns, means of three experiments, with triplicates of each experiment; bars, SD. *, p <0.05, significantly different from non treated control group. #, p <0.01 significantly different from SVT-treated group.

Article Snippet: Small interfering (si) RNA species for death receptor (DR4 and DR5) and non-targeting control siRNA were purchased from Bioneer (Daejeon, Korea), and death receptor 4 (DR4) was purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA)

Techniques: Expressing, Western Blot, Control

Journal: BMC Cancer

Article Title: Snake venom toxin from vipera lebetina turanica induces apoptosis of colon cancer cells via upregulation of ROS- and JNK-mediated death receptor expression

doi: 10.1186/1471-2407-12-228

Figure Lengend Snippet: Effect of ROS on upregulation of DR4 or DR5 through JNK activation by snake venom toxin. a , Effect of antioxidant (NAC) on the cell viabilty induced by snake venom toxin. Cells were pretreated with various concentratins of NAC (0, 1, 10 mM) for 1 h and then treated with 1 μg/ml of snake venom toxin for 30 min, and whole cell extracts were analyzed by western blotting using the relevant antibodies. b , Effect of NAC on the expression of death receptors and JNK phosphorylation. Cells were pretreated with NAC for 1 h, and then cells were treated with snake venom toxin for 30 min, and whole cell extracts were analyzed by Western blotting using DR4, DR5, p-JNK and β-actin antibodies. Each band is representative for three experiments. Columns, means of three experiments, with triplicates of each experiment; bars, SD. *, p <0.05, significantly different from non treated control group. #, p <0.01 significantly different from SVT-treated group.

Article Snippet: Small interfering (si) RNA species for death receptor (DR4 and DR5) and non-targeting control siRNA were purchased from Bioneer (Daejeon, Korea), and death receptor 4 (DR4) was purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA)

Techniques: Activation Assay, Western Blot, Expressing, Phospho-proteomics, Control

Journal: Nature cell biology

Article Title: Loss of the fragile X syndrome protein FMRP results in misregulation of nonsense-mediated mRNA decay

doi: 10.1038/s41556-020-00618-1

Figure Lengend Snippet: a, Quantitations demonstrating that NMD was inhibited by each of the 3 NMD inhibitors. Samples derived from day 0, that is 24-hr after culturing iPSCs in the presence of each inhibitor as shown in Fig. 4e, b, Results are means with S.D., where n = 3 independent biological replicates. (*) P < 0.05 or (**) P < 0.01 is relative to RNA samples without NMD inhibitor (two-sided t-test). b, As in Extended Data Fig. 7j, but with or without an NMD inhibitor. Means with S.D., where n = 5 independent biological replicates. (*) P < 0.05, (**) P < 0.01 or (***) P < 0.01 is relative to RNA samples without an NMD inhibitor (two-sided t-test). c, As in Fig. 4e, but staining for MAP2 (red). d, As in Fig. 4e, but staining for BRN2/POU3F2 (green). Results are representative of 3 independent biological replicates in (c) and (d). e, Histogram representations of quantitations of western blots shown in Fig. 4f. Results represent n = 3 independent biological replicates, except for SYN1 analyses in FXS neurons, BRN2 analysis in FXS neurons treated with NMDI-1, and DCX analysis in FXS neurons treated with curcumin, where n = 2 independent biological replicates. f, Histogram representation of neurite outgrowth manifested by representative normal or FXS neurons on day 15 after differentiation. Results represent 2 independent biological replicates for FXS neurons and 3 independent biological replicates for normal neurons. All cells were stained for viability, and fluorescent signals derive from minimally 4 fields per well, where the extent of fluorescence for FXS neurons in the absence (−) of inhibitor is defined as 1. Statistical source data are provided in Source Data Extended Data Fig. 8.

Article Snippet: Coverslips were blocked with 3% bovine serum albumin (Rockland Immunochemicals) in Tris-buffered saline containing 0.1% Tween 20 (TBS-T) for 30 min at room temperature, washed once with TBS-T and incubated overnight at 4 °C in primary antibody that had been diluted in TBS-T using the following antibodies (see Supplementary Table 6 ): anti-p-UPF1 S1116 (1:250), anti-p-UPF1 S1089 (1:250), anti-UPF1 (1:500), anti-FMRP (1:250), anti-TRA-1–60 (1:500), anti-OCT4 (1:500), anti-MAP2 (1/250), anti-β3-Tubulin/TUJ1 (1:500) and anti-BRN2/POU3F2 (1:250).

Techniques: Derivative Assay, Staining, Western Blot, Fluorescence