Fig 2: The effect of ISL on the levels of anti-apoptotic and pro-apoptotic proteins in IL-1β-induced chondrocyte-like ATDC5 cells. (A) The IL-1β stimulated the cells were treated with different doses of ISL (2.5, 5 and 10 µmol/l) for 48 h. Then protein expressions Level of Bcl-2, Bax, caspase-3, cleaved-caspase-3, caspase-9 and cleaved-caspase-9 were detected by western blotting. (B) The protein analysis of quantitative fluorescence. GAPDH was used as internal control. The values represent the mean ± standard deviation of three independent experiments. *P<0.05 vs. model group treated with only IL-1β. ISL, isoliquiritigenin; IL, interleukin.

Fig 3: Downregulation of BPTF sensitized chemotherapeutic drugs-mediated anti-neoplastic effects in HCC cells. (A) Hep3B and HepG2 cells with stable knockdown of BPTF were treated with different concentrations of 5-FU or DDP and the cell viability was tested by MTT assay. (B) The IC50 value of 5-FU and DDP was calculated on the basis of Fig. 4A and shown. (C) Acridine orange/ethidium bromide double staining method was used to evaluate the degree of apoptosis caused by DDP upon BPTF silencing and the pictures were taken by Inverted fluorescence microscope. Scale bars, 20 µm. (D) FACS analysis was used to detect the apoptosis by FITC-AV/PI staining in HepG2 and Bel7402 cells mediated by DDP upon BPTF silencing. (E) The expression level of cleaved-caspase-7, cleaved-caspase-9, cleaved-PARP and Bcl-2 was determined by western blot in HepG2 and Bel7402 cells treated by DDP upon BPTF silencing.

Fig 4: miR-125b promotes DOX-induced cell death through caspase-dependent apoptosis in MCF-7/R cells. (A) MCF-7/R cells were transfected with RNAs and plasmids for 24 h. Then, the cells were treated with 4 µg/ml DOX for another 48 h. Annexin V/PI staining was performed to detect the apoptosis rate in MCF-7/R cells. *P<0.05 vs. DOX+miR-NC group; #P<0.05 vs. DOX+miR-125b group. (B) Western blot analysis was performed to measure the cleavage of caspase-9, caspase-3 and its substrate PARP. DOX, doxorubicin; miR, microRNA; MCF-7/R, DOX-resistant MCF-7 cells; PI, propidium iodide; HAX-1, hematopoietic cell-specific protein 1-associated protein X-1; NC, negative control; PARP, poly(ADP-ribose)polymerase.

Fig 5: Effects of PADI2 knockout on the relative expression levels of the activated caspase-3, -2, -8, and -9. In the differentiated macrophages stimulated with LPS, the protein expression levels of the intact caspases and the cleaved caspases were performed by Western blotting in the PADI2 knockout group and the control group. The relative protein expression was defined as the ratio of intact caspases or cleaved caspases to the actin band intensity: (A) caspase-3; (B) caspase-2; (C) caspase-8; (D) caspase-9.