Fig. 5: USP7 stabilizes NLRP3 via decreasing K48-linked ubiquitination of NLRP3.

A The SDS-PAGE band pattern of immunoprecipitated proteins using anti-USP7 and IgG. The bands of protein were excised to mass spectrometry (MS) analysis for identification. The red arrow marks the distinct band (NLRP3) in USP7 immunoprecipitated samples. B The immunoprecipitated protein of NLRP3 were analyzed using anti-USP7 and IgG. C The protein expression levels of NLRP3, pro-caspase1, and caspase1 in MG63 and U2OS cells treated with 100 nM of E2 for 48 h with or without pretreatment of 1 μM of torin1. D The protein expression levels of NLRP3 and USP7 in U2OS cells treated with shCtrl or shUSP7 for 24 h. E U2OS cells were transfected with shCtrl or shUSP7 for 24 h. The protein extracts were immunoprecipitated with IgG beads of anti-NLRP3, and the protein expression levels of K48-ubiquitin, K63-ubiquitin and NLRP3 were detected. F The relative expression of NLRP3 in OS tissues and normal tissues (n = 8). G The correlation between USP7 and NLRP3 in OS tissues or normal tissues (n = 8). H The colabel staining of USP7 and NLRP3 in OS tissues or normal tissues, scale bar: 50 µm. Arrow indicates USP7⁺NLRP3⁺ cells. I Quantification of the USP7⁺NLRP3⁺ cells in OS tissues or normal tissues (n = 8). **P < 0.01, ****P < 0.0001.