Fig 2: Compound 8a sensitizes pancreatic cancer cells to gemcitabine.a Cell viability of BXPC-3 cells after exposure to increasing concentration of gemcitabine in presence or absence of 6.25 μM 8a for 48 h. b CompuSyn software calculated viability data and plotted Fa-CI curves to analyze the synergistic effects of the two drugs. c Cell viability of HUVEC cells after exposure to increasing concentration of gemcitabine in presence or absence of 6.25 μM 8a for 48 h. d The colony formation assay of BXPC-3 cells after treatment with 10 nM gemcitabine, 6.25 μM 8a, or their combination for 10 days. e Western blotting analysis of proliferation-related protein expression. f Analysis of apoptosis by the Muse cell analyzer in BXPC-3 cells following exposure to gemcitabine or/and 8a. g Western blot analysis of cleaved-PARP, cleaved-Caspase 3, and cleaved-Caspase 9 in BXPC-3 cells after indicated treatment. Data are presented as mean ± SD (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, versus control. ###P < 0.001, versus GEM group.

Fig 3: PINK1/Parkin-dependent mitophagy decreases mitochondrial ROS production and apoptosis induced by Stx2A. (A, B) Caco-2 cells were cotransfected with 1 μg/ml Stx2A plasmids and control siRNA, PINK1 siRNA, or Parkin siRNA. Cells were then treated with MitoTEMPO (100 μM) for 4 h followed by additional culture for 24 h. Representative images of mitochondrial ROS (MitoSOX, Red) and 8-OHdG (Green) immunofluorescence (A) as well as flow cytometry analysis of mitochondrial ROS by MitoSOX (B) in Caco-2 cells. Scale bar = 5 μm. (C) Representative Western blot analysis of PINK1, Parkin, LC3 II/I, Beclin1, Tom20 and SQSTM1 after transfection with 1 μg/ml Stx2A plasmids for 24 h followed by treatment with MitoTEMPO (100 μM) for 4 h. (D, E) Caco-2 cells were treated with or without MitoTEMPO after silencing PINK1 or Parkin and transfection with Stx2A plasmids. Western blot analysis (D) of Cleaved-Caspase-3, Cleaved-Caspase-9, Bax and Bcl-2. Representative images and quantification of cell apoptosis by flow cytometry (E). n = 3. *P < 0.05.

Fig 4: Metapristone inhibited endometrial cancer cell growth and promoted endometrial cancer cell apoptosis. a Cell growth curves by MTT assay of RL95-2 cells and Ishikawa cells with metapristone (50 μM) treatment. Ethanol-treated cells were as control. Results represent mean ± SD (n = 6). b Cell colony formation of RL95-2 cells and Ishikawa cells with metapristone (50 μM) treatment. Ethanol-treated cells were as control. Values are mean ± SD (n = 3 per group). c EdU assay was used to detect the cell proliferation phenotype with the treatment of ethanol or metapristone (50 μM 48 h). d The protein level of Cleaved-Caspase 3, Cleaved-Caspase 9, Bax and Bcl-2 in RL95-2 cells and Ishikawa cells with ethanol or metapristone treatment were detected by western blot. GAPDH was used as the control

Fig 5: UPM-induced apoptosis through a caspase-dependent pathway. (A) Increased TUNEL assay staining and (B) cell counts revealed an increase of apoptosis cells in MGs of UPM-treated mice compared with those of the control. (C) Immunohistochemical staining of cell apoptosis marker cleaved caspase-3 revealed increased expression level in acini in UPM-treated mice. (D–G) The expression of apoptosis-related genes showed an obvious elevation of (E) cleaved caspase-8, (F) cleaved caspase-9, and (G) cleaved caspase-3 in UPM-treated mice (all n = 4 mice per group). Scale bars, 50 µm (A) and 20 µm (C). Data are shown as mean ± SD. *p < 0.05, **p < 0.01 and ***p < 0.001.