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. 2021 Nov 11;2021:1806344. doi: 10.1155/2021/1806344

Figure 7.

Figure 7

PFKFB3 modulated the NF-κB signaling as well as the NLRP3 inflammasome-induced pyroptosis while the pharmacological concentration of MET suppressed pyroptosis partly via inhibiting PFKFB3. HTR-8/SVneo cells were transfected with PFKFB3 plasmid for overexpression or negative vector; 200 ng/ml LPS, 10 μM MET, and 10 μM 3PO were subsequently incubated. (a–k) Western blot analysis of PFKFB3, NF-κB1, p65, p-p65, IκB, p-IκB, p-IKK, NLRP3, caspase1-p10, GSDMD, and ASC protein expression and densitometry quantification of PFKFB3 (b), NF-κB1 (c), p-p65/p65 (d), IκB (e), p-IκB (f), p-IKK (g), NLRP3 (h), caspase1-p10 (i), GSDMD (j), ASC (k) levels. (l) Analysis of NLRP3 ubiquitination levels in HTR-8/SVneo cells of different groups. (m, n) ELISA analysis of IL-1β (m) and IL-18 (n) concentrations in cell culture medium. (o, p) Representative immunofluorescence images and quantification of double-fluorescent staining with PI (red) and Hoechst33342 (blue). Scale bar: 100 μm. Data are shown as the mean ± SD from three independent experiments. ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001 by Student's t-test. SD: standard deviation; NC-OE: negative vector; PFKFB3-OE: PFKFB3 overexpression plasmid.