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. 2022 Mar 12;8:112. doi: 10.1038/s41420-022-00906-9

Fig. 2. TNF-α induces pyroptosis and upregulates HDAC11 expression in HUVECs.

Fig. 2

A HUVECs were treated with TNF-α (10, 20, 40, 60, or 80 ng/mL) for 12 h. GSDMD, GSDMD-N, GSDME, and GSDME-N protein expressions were determined by Western blotting. B HUVECs were treated with 40 ng/mL TNF-α for 2, 4, 8, 12, or 24 h. GSDMD, GSDMD-N, GSDME, and GSDME-N protein expressions were determined by Western blotting. C The levels of IL-1β, IL-6, and IL-18 in the cellular supernatant were detected by ELISA assay. D The cellular supernatant LDH level was evaluated with a cytotoxicity detection LDH kit. E The representative photographs of double-fluorescent staining with PI (red) and Hoechst 33342 (blue), and the quantification of PI-positive cells, ×200. Scale bar indicates 100 μm. F Immunofluorescence staining was performed to determine the expression and localization of HDAC11, ×200. Scale bar indicates 50 μm. G, H The protein and mRNA expressions of HDAC11 were determined by Western blotting and quantitative real-time PCR. *P < 0.05, **P < 0.01 vs. control group. Results are expressed as mean ± SD (n = 3).