Journal: Cell Death Discovery

Article Title: Verteporfin reverses progestin resistance through YAP/TAZ-PI3K-Akt pathway in endometrial carcinoma

doi: 10.1038/s41420-023-01319-y

Figure Lengend Snippet: A , B The inhibition of YAP/TAZ by Verteporfin in dose-dependent and time-dependent manners by western blotting assay. C The growth curves of IshikawaPR cells after treated with medium, MPA, Verteporfin, or MPA plus Verteporfin. ** P < 0.01 IshikawaPR cells treated with medium vs IshikawaPR cells treated with MPA plus Verteporfin. D The colony formation ability of IshikawaPR cells with treatment of medium, MPA, Verteporfin, and MPA plus Verteporfin. E The apoptosis of IshikawaPR cells with treatment of medium, MPA, Verteporfin or MPA plus Verteporfin detected by flow cytometry assay. F The effects of Verteporfin, MPA, and MPA plus Verteporfin on tumors growth in vivo. G Representative images of IHC staining of YAP, TAZ, Ki67, cleaved caspase 3, and p-Akt in tumor tissues. * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001.

Article Snippet: Antibodies for YAP1(ab76252), PGR(ab23085), and TBP (ab818) were purchased from Abcam (Cambridge, UK); TAZ (#83669), β-actin(#4970), MST1(#3682), parp(#2772), cleaved caspase 3(#9664), cleaved caspase 7(#8438), cleaved caspase 9(#20750), bax(#2772), and CyclinD1(#2978), Akt(#4691), p-Akt(#13038), anti-flag(#14793), anti-rabbit IgG(#7074), anti-mouse IgG(#7076) were from Cell Signaling Technology (Danvers, MA,USA).

Techniques: Inhibition, Western Blot, Flow Cytometry, In Vivo, Immunohistochemistry

Journal: Frontiers in Oncology

Article Title: Suppression of head and neck cancer cell survival and cisplatin resistance by GRP78 small molecule inhibitor YUM70

doi: 10.3389/fonc.2022.1044699

Figure Lengend Snippet: YUM70 reduces cell viability and restores cisplatin sensitivity in cisplatin-resistant SCC15. (A) Cisplatin-resistant SCC15 cells (the cisR-3 line) were treated with increasing doses of YUM70 for 48 hr. Cell viability was determined by WST-1 assay. DMSO was used as vehicle control. (B) Representative images of cisR-3 cells treated with the indicated dosages of YUM70 alone or in combination with 12 μM cisplatin for 48 hr. Scale bar = 100 μm. (C) Western blot of lysates of cisR-3 cells treated with the indicated dosages of YUM70 for 48 hr, alone or in combination with 12 μM cisplatin. The protein levels of cleaved Caspase 7 (c-Cas7) and cleaved-PARP (c-PARP) were measured, with GAPDH serving as loading control. (D) Cell viability of cisR-3 cells treated with the indicated dosages of YUM70, alone or in combination with the indicated dosages of cisplatin for 48 hr was measured by WST-1 assay. Data are presented as means ± S.D. (n=3).

Article Snippet: Primary antibodies used in this study were as follows: mouse anti-GRP78 (BiP, 1:2000, BD Transduction Laboratories #610979), mouse anti-GAPDH (1:2000, Santa Cruz #sc-32233), mouse anti-CHOP (1:1000, Cell Signaling #2895), rabbit anti-cleaved-PARP (1:1000, Cell Signaling #5625), and rabbit anti-cleaved-Caspase 7 (1:1000, Cell Signaling #8438).

Techniques: WST-1 Assay, Western Blot

Journal: Frontiers in Oncology

Article Title: Suppression of head and neck cancer cell survival and cisplatin resistance by GRP78 small molecule inhibitor YUM70

doi: 10.3389/fonc.2022.1044699

Figure Lengend Snippet: YUM70 treatment induces apoptosis in cisplatin-treated cisR-3 spheroids. (A) A representative image of cisR-3 spheroids. Scale bar = 200 μm. (B) GRP78 protein levels in cisR-3 grown in 2D culture and 3D spheroid culture was determined by Western blot with GAPDH serving as loading control. (C) Quantitative results of (B) . (D) Western blot analysis of lysates of cisR-3 spheroids treated with the indicated dosages of cisplatin for 48 hr, alone or in combination with 10 μM YUM70. The protein levels of GRP78, CHOP, cleaved Caspase 7 (c-Cas7), and cleaved-PARP (c-PARP) were determined with GAPDH serving as loading control. (E-G) Quantitative results of (D) after normalization against the GADPH level. Data are presented as means ± S.D. (n=3). * P ≤ 0.05, ** P ≤ 0.01, ns, not significant (Student’s t test).

Article Snippet: Primary antibodies used in this study were as follows: mouse anti-GRP78 (BiP, 1:2000, BD Transduction Laboratories #610979), mouse anti-GAPDH (1:2000, Santa Cruz #sc-32233), mouse anti-CHOP (1:1000, Cell Signaling #2895), rabbit anti-cleaved-PARP (1:1000, Cell Signaling #5625), and rabbit anti-cleaved-Caspase 7 (1:1000, Cell Signaling #8438).

Techniques: Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Protective Effects of Hexarelin and JMV2894 in a Human Neuroblastoma Cell Line Expressing the SOD1-G93A Mutated Protein

doi: 10.3390/ijms24020993

Figure Lengend Snippet: Caspase-3 and caspase-7 mRNA levels following incubation with increasing concentrations of H 2 O 2 . SH-SY5Y WT and SOD1 G93A cells were incubated with different concentrations of H 2 O 2 alone (0, 100, 150, 200 μM). Caspase-3 ( A ) and caspase-7 ( B ) mRNA levels were measured by RT-PCR and normalized for the respective β-actin mRNA levels. Data are expressed as mean ± SEM of 18 replicates obtained in 3 independent experiments ( n = 6 in each experiment). For SH-SY5Y WT: § p < 0.05 vs. CTRL; for SH-SY5Y SOD1 G93A * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. CTRL.

Article Snippet: After 3 washes in PBS-T, membranes were incubated with the primary antibody overnight at 4 °C (Anti-cleaved caspase-3 (Asp175) (5A1E) rabbit antibody, #9664, Cell Signaling Technology, 1:1000; anti-cleaved caspase-7 (Asp198) rabbit antibody, #9491, Cell Signaling Technology, 1:1000; anti-Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) rabbit antibody, #9101, Cell Signaling Technology, 1:1000; anti-p44/42 MAPK (Erk1/2) rabbit antibody, #4695, Cell Signaling Technology, 1:1000; anti-Phospho-p38 MAPK (Thr180/Tyr182) rabbit antibody, #4511, Cell Signaling Technology, 1:1000; anti-p38 MAPK rabbit antibody, #9212, Cell Signaling Technology, 1:1000; anti-Phospho-Akt rabbit antibody, #4060, Cell Signaling Technology, 1:2000; anti-Akt rabbit antibody, #4685, Cell Signaling Technology, 1:1000; anti-actin rabbit antibody, #A2066, Sigma Aldrich, 1:2500), and then with a peroxidase-coupled goat anti-rabbit IgG (#31460, Thermo Scientific, 1:5000) for 1 h at room temperature.

Techniques: Incubation, Reverse Transcription Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: Protective Effects of Hexarelin and JMV2894 in a Human Neuroblastoma Cell Line Expressing the SOD1-G93A Mutated Protein

doi: 10.3390/ijms24020993

Figure Lengend Snippet: Caspase-3 and caspase-7 mRNA levels following incubation with H 2 O 2 and GHS. The SH-SY5Y SOD1 G93A were also co-incubated with 1 µM GHS and 150 µM H 2 O 2 for 24 h. Caspase-3 ( A ) and caspase-7 ( B ). The mRNA levels were measured by RT–PCR and normalized for the respective β-actin mRNA levels. Data are expressed as mean ± SEM of 18 replicates obtained in 3 independent experiments ( n = 6 in each experiment). * p < 0.05, and *** p < 0.001 vs. CTRL; ° p < 0.05 vs. H 2 O 2 .

Article Snippet: After 3 washes in PBS-T, membranes were incubated with the primary antibody overnight at 4 °C (Anti-cleaved caspase-3 (Asp175) (5A1E) rabbit antibody, #9664, Cell Signaling Technology, 1:1000; anti-cleaved caspase-7 (Asp198) rabbit antibody, #9491, Cell Signaling Technology, 1:1000; anti-Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) rabbit antibody, #9101, Cell Signaling Technology, 1:1000; anti-p44/42 MAPK (Erk1/2) rabbit antibody, #4695, Cell Signaling Technology, 1:1000; anti-Phospho-p38 MAPK (Thr180/Tyr182) rabbit antibody, #4511, Cell Signaling Technology, 1:1000; anti-p38 MAPK rabbit antibody, #9212, Cell Signaling Technology, 1:1000; anti-Phospho-Akt rabbit antibody, #4060, Cell Signaling Technology, 1:2000; anti-Akt rabbit antibody, #4685, Cell Signaling Technology, 1:1000; anti-actin rabbit antibody, #A2066, Sigma Aldrich, 1:2500), and then with a peroxidase-coupled goat anti-rabbit IgG (#31460, Thermo Scientific, 1:5000) for 1 h at room temperature.

Techniques: Incubation, Reverse Transcription Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: Protective Effects of Hexarelin and JMV2894 in a Human Neuroblastoma Cell Line Expressing the SOD1-G93A Mutated Protein

doi: 10.3390/ijms24020993

Figure Lengend Snippet: GHS inhibits apoptotic pathway through caspases inactivation. The SH-SY5Y SOD1 G93A cells were treated with H 2 O 2 alone, GHS alone, or with a combination of GHS and H 2 O 2 for 24 h. Western blot assays were used to measure levels of ( A ) cleaved caspase-3/β-actin, and ( B ) cleaved caspase-7/β-actin. All assays were performed in at least 3 independent experiments ( n = 3). Statistical significance: ** p < 0.01, and *** p < 0.001 vs. CTRL; °° p < 0.01 vs. H 2 O 2 .

Article Snippet: After 3 washes in PBS-T, membranes were incubated with the primary antibody overnight at 4 °C (Anti-cleaved caspase-3 (Asp175) (5A1E) rabbit antibody, #9664, Cell Signaling Technology, 1:1000; anti-cleaved caspase-7 (Asp198) rabbit antibody, #9491, Cell Signaling Technology, 1:1000; anti-Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) rabbit antibody, #9101, Cell Signaling Technology, 1:1000; anti-p44/42 MAPK (Erk1/2) rabbit antibody, #4695, Cell Signaling Technology, 1:1000; anti-Phospho-p38 MAPK (Thr180/Tyr182) rabbit antibody, #4511, Cell Signaling Technology, 1:1000; anti-p38 MAPK rabbit antibody, #9212, Cell Signaling Technology, 1:1000; anti-Phospho-Akt rabbit antibody, #4060, Cell Signaling Technology, 1:2000; anti-Akt rabbit antibody, #4685, Cell Signaling Technology, 1:1000; anti-actin rabbit antibody, #A2066, Sigma Aldrich, 1:2500), and then with a peroxidase-coupled goat anti-rabbit IgG (#31460, Thermo Scientific, 1:5000) for 1 h at room temperature.

Techniques: Western Blot