Fig. 3: IL-1β^low enhances resilience to cytokine-induced death in INS-1E cells.

A, B INS-1E cells were conditioned with IL-1β^low and subsequently challenged or not with CYT. After indicated time, cell death was analyzed by fluorescence microscopy after Hoechst (blue)/propidium iodide (red) dual staining. A Representative images of cells under indicated experimental conditions; scale bars 10 μm. Selected apoptotic cells are shown at higher magnification in the inset, n = 4. B Percentage of apoptotic cells based on analysis of ten high-power fields per condition, n = 4. C, D IL-1β^low-preconditioned INS-1E cells were challenged or not with CYT for 48 h. Apoptosis was quantified by flow cytometry after Annexin-V/7AAD dual staining. C Representative dot plots of cells under each experimental condition. D Percentage of early (Q3) and late (Q2) apoptotic cells, n = 3. E–J IL-1β^low-preconditioned INS-1E cells were treated or not with CYT for 16 h. E DP5 mRNA levels, FPUMA mRNA levels, and GBax/Bcl-2 mRNA ratio was analyzed by RT-qPCR. Relative mRNA levels were normalized to HPRT, n = 3. H–J CHOP and cleaved caspase-3 protein levels were analyzed by Western blot; β-actin was used as loading control, n = 3. Data are shown as mean ± SD. (*) p < 0.05, (**) p < 0.01.