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(A) Structure of the IRS reporter. The Cal27-ATF4-GFP reporter cells were treated with PG3, cisplatin, and <t>thapsigargin</t> (TG) for 4 or 8 hours, respectively. Western blot was performed using the indicated antibodies. (B) Flow cytometric analysis of cell death. Cal27 cells were pre-treated with 2Bact (20 µM) for 30 min, then PG3 (0.5 µM), and cisplatin (5 µg/mL) were added to corresponding wells, and incubated at 37°C for 12 hours. (C) and (D) Cells were pre-treated with z-VAD-FMK for 30 min, then were treated with the combinations of PG3 plus cisplatin for 16 hours (C) , and 21 hours (D) , respectively. Western blots were performed using the indicated antibodies. (E) and (F) Cal27 and FaDu cells were pre-treated with 2Bact (20 µM) for 30 min, then the combined treatments of PG3 plus cisplatin were performed for Cal27 (16 hours), and FaDu (21 hours). Western blot assays were performed using the indicated antibodies.
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(A) Structure of the IRS reporter. The Cal27-ATF4-GFP reporter cells were treated with PG3, cisplatin, and <t>thapsigargin</t> (TG) for 4 or 8 hours, respectively. Western blot was performed using the indicated antibodies. (B) Flow cytometric analysis of cell death. Cal27 cells were pre-treated with 2Bact (20 µM) for 30 min, then PG3 (0.5 µM), and cisplatin (5 µg/mL) were added to corresponding wells, and incubated at 37°C for 12 hours. (C) and (D) Cells were pre-treated with z-VAD-FMK for 30 min, then were treated with the combinations of PG3 plus cisplatin for 16 hours (C) , and 21 hours (D) , respectively. Western blots were performed using the indicated antibodies. (E) and (F) Cal27 and FaDu cells were pre-treated with 2Bact (20 µM) for 30 min, then the combined treatments of PG3 plus cisplatin were performed for Cal27 (16 hours), and FaDu (21 hours). Western blot assays were performed using the indicated antibodies.
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(A) Structure of the IRS reporter. The Cal27-ATF4-GFP reporter cells were treated with PG3, cisplatin, and thapsigargin (TG) for 4 or 8 hours, respectively. Western blot was performed using the indicated antibodies. (B) Flow cytometric analysis of cell death. Cal27 cells were pre-treated with 2Bact (20 µM) for 30 min, then PG3 (0.5 µM), and cisplatin (5 µg/mL) were added to corresponding wells, and incubated at 37°C for 12 hours. (C) and (D) Cells were pre-treated with z-VAD-FMK for 30 min, then were treated with the combinations of PG3 plus cisplatin for 16 hours (C) , and 21 hours (D) , respectively. Western blots were performed using the indicated antibodies. (E) and (F) Cal27 and FaDu cells were pre-treated with 2Bact (20 µM) for 30 min, then the combined treatments of PG3 plus cisplatin were performed for Cal27 (16 hours), and FaDu (21 hours). Western blot assays were performed using the indicated antibodies.

Journal: bioRxiv

Article Title: Integrated stress response activation in combination PG3 and cisplatin-treated TP53 -mutated head and neck squamous carcinoma cells induces potent apoptotic response

doi: 10.64898/2026.09.22.752907

Figure Lengend Snippet: (A) Structure of the IRS reporter. The Cal27-ATF4-GFP reporter cells were treated with PG3, cisplatin, and thapsigargin (TG) for 4 or 8 hours, respectively. Western blot was performed using the indicated antibodies. (B) Flow cytometric analysis of cell death. Cal27 cells were pre-treated with 2Bact (20 µM) for 30 min, then PG3 (0.5 µM), and cisplatin (5 µg/mL) were added to corresponding wells, and incubated at 37°C for 12 hours. (C) and (D) Cells were pre-treated with z-VAD-FMK for 30 min, then were treated with the combinations of PG3 plus cisplatin for 16 hours (C) , and 21 hours (D) , respectively. Western blots were performed using the indicated antibodies. (E) and (F) Cal27 and FaDu cells were pre-treated with 2Bact (20 µM) for 30 min, then the combined treatments of PG3 plus cisplatin were performed for Cal27 (16 hours), and FaDu (21 hours). Western blot assays were performed using the indicated antibodies.

Article Snippet: Cisplatin, 2Bact, silmitasertib, and pevonedistat (MedChemExpress); Z-VAD-FMK, MG-132, and SB216763 (Selleckchem); Thapsigargin (Tocris Bioscience); D4476 (Cayman Chemicals), and ONC201 (Chimerix, Durham, NC).

Techniques: Western Blot, Analysis, Incubation

(A) Cal27-ATF4-GFP reporter cells were treated with TG or ONC201 for 24 hours. Images were taken using the ImageXpress® Micro Confocal High-Content Imaging system. (B) FaDu-ATF4-GFP reporter cells were treated with PG3, cisplatin, and thapsigargin (TG) for 6 or 16 hours, respectively. Western blot was performed using the indicated antibodies.

Journal: bioRxiv

Article Title: Integrated stress response activation in combination PG3 and cisplatin-treated TP53 -mutated head and neck squamous carcinoma cells induces potent apoptotic response

doi: 10.64898/2026.09.22.752907

Figure Lengend Snippet: (A) Cal27-ATF4-GFP reporter cells were treated with TG or ONC201 for 24 hours. Images were taken using the ImageXpress® Micro Confocal High-Content Imaging system. (B) FaDu-ATF4-GFP reporter cells were treated with PG3, cisplatin, and thapsigargin (TG) for 6 or 16 hours, respectively. Western blot was performed using the indicated antibodies.

Article Snippet: Cisplatin, 2Bact, silmitasertib, and pevonedistat (MedChemExpress); Z-VAD-FMK, MG-132, and SB216763 (Selleckchem); Thapsigargin (Tocris Bioscience); D4476 (Cayman Chemicals), and ONC201 (Chimerix, Durham, NC).

Techniques: Confocal, Imaging, Western Blot