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

Fig. 3. Blockade of TNFR1 inhibits pyroptosis and downregulates HDAC11 expression in TNF-α-induced HUVECs.

Fig. 3

HUVECs were treated with TNF-α (20, 40, or 80 ng/mL) for 12 h. A, B The expressions of TNFR1 and TNFR2 protein and mRNA were determined by Western blotting and quantitative real-time PCR, respectively. The HUVECs were pretreated with a TNFR1 neutralizing antibody (5 µg/mL) for 30 min and then stimulated with TNF-α (40 ng/mL) for 12 h. C The protein expressions of GSDMD, GSDMD-N, GSDME, and GSDME-N were determined by Western blotting. D The levels of IL-1β, IL-6, and IL-18 in the cellular supernatant were detected by ELISA assay. E The cellular supernatant LDH level was evaluated with a cytotoxicity detection LDH kit. F The representative photographs of double-fluorescent staining with PI (red) and Hoechst 33342 (blue), and the quantification of PI-positive cells, ×100. Scale bar indicates 200 μm. G Immunofluorescence staining was performed to determine the expression and localization of HDAC11, ×200. Scale bar indicates 50 μm. H, I The protein and mRNA expressions of HDAC11 were determined by Western blotting and quantitative real-time PCR. **P < 0.01, vs. NC siRNA group; #P < 0.05, ##P < 0.01 vs. NC siRNA + TNF-α group. Results are expressed as mean ± SD (n = 3).