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. 2024 Apr 10;15(4):255. doi: 10.1038/s41419-024-06631-8

Fig. 4. BZA-dependent inhibition of gp130/STAT3 signalling sensitises colorectal cancer cells to SMAC-mimetic-induced apoptotic cell death.

Fig. 4

A Western blot of cIAP-1, cIAP-2, XIAP and GAPDH protein expression in LIM2405 cells treated with 15 μM BZA, 5 μM LCL161, 5 μM BPT (Birinapant) alone and 15 μM BZA + 5 μM LCL161, 15 μM BZA + 5 μM BPT in the presence of 30 ng/ml IL-11 and 10 ng/ml TNF-α for 24 hours. B Flow cytometry analysis of Annexin V-FITC and TO-PRO-3-iodide stained LIM2405 cells treated with 15 μM BZA, 5 μM LCL161, 5 μM BPT (Birinapant) alone and 15 μM BZA + 5 μM LCL161, 15 μM BZA + 5 μM BPT in the presence of 30 ng/ml IL-11 and 10 ng/ml TNF-α for 8 hours. Error bars are mean ± SEM of n = 3 replicates. Student’s unpaired t-test: ***p < 0.001, **p < 0.01. C, D % Cell viability of LIM2405 cells treated with the indicated concentrations of (C) BZA + LCL161 and (D) BZA + BPT in the presence of 30 ng/ml IL-11 and 10 ng/ml TNF-α for 8 hours compared to the untreated control. Data is representative of the mean of n = 3 replicates. Statistical analysis was conducted using a two way ANOVA with Tukeys multiple comparisons test. Interaction factor between BZA and LCL161 p < 0.001. Interaction factor between BZA and BPT p < 0.05. (E) Schematic representation of the proposed mechanism of action of BZA combined with SMAC-mimetics induce tumour cell apoptosis.