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. 2021 Sep 11;29(2):439–450. doi: 10.1038/s41418-021-00867-z

Fig. 5. Depletion of TRIM21 reduces GSDMD-N upon inflammatory stimulation.

Fig. 5

a WT, Gsdmd−/−, Trim21−/−, and Gsdmd−/− Trim21−/− iBMDM cells were with 1 μg/mL LPS for 2 h followed by 20 μM Nig. Gd−/− T21−/− denotes double knockout Gsdmd−/− Trim21−/− . Cell death was measured by the LDH Cytotoxicity Assay Kit (Promega). The data are the means ± SD of triplicate samples from a representative experiment (****P < 0.0001; ns no significance). b–e Immunoblotting analysis of the protein expression levels (TRIM21, GSDMD, NLRP3, Caspase-1, p-RIP3, and Caspase-3) in WT, Gsdmd−/− , Trim21−/−, and Gsdmd−/− Trim21−/− iBMDM cell lines treated with (e) or without (b) LPS plus Nig as described in (a). Quantifications of the results for GSDMD, GSDMD-N, Caspase-3, and p-RIP3 were shown in (c) (related to b) and (d) (related to e). The soluble part contained cytoplasmic proteins. The insoluble part contained proteins in the membranes, cell organelles, and nuclei. All the proteins were extracted by the Soluble & Insoluble Protein Extraction Kit (Sangon Biotech, China). p50, pro-caspase-1; p10 and p25, cleaved-caspase-1. p17, cleaved-caspase-3. All data are representative of three independent experiments.