Fig. 6: GSDME promotes translocation of p65 and c-jun into nucleus to upregulate psoriatic inflammation. | Cell Death & Disease

Fig. 6: GSDME promotes translocation of p65 and c-jun into nucleus to upregulate psoriatic inflammation.

From: Gasdermin E promotes translocation of p65 and c-jun into nucleus in keratinocytes for progression of psoriatic skin inflammation

Fig. 6Fig. 6

A Proteins of GSDME FL and GSDME NT were detected by western blotting in HaCaT cells at 12, 24, and 48 h after treatment with or without M5. Statistical analysis of interested protein levels was shown. B HaCaT cells were transfected with lentivirus vector containing either NC shRNA or GSDME shRNA, establishing GSDME knockdown cells (sh GSDME) and control cells (sh NC). Knockdown efficiency was evaluated using western blotting assay. Statistical analysis of GSDME was shown. C, D Immunofluorescence assay was used to detect Ki-67 expression or Edu-positive cells. Statistical analysis of mean immunofluorescence intensity was shown. E Quantitative real-time PCR analysis was performed to determine mRNA levels of Il1b, Ccl20, Cxcl1, Cxcl8, S100a8, and S100a9. Statistical analysis was shown. F Proteins of GSDME FL, S100A8, and S100A9 in cell lysate were detected by western blotting assay. Statistical analysis of interested protein levels was shown. G Secretion levels of S100A8/A9, CCL20, and CXCL8 were determined by ELISA assay. Statistical analysis was shown. H S100A8 and S100A9 were detected by immunofluorescence assay in mice skin sections, respectively. Statistical analysis was shown. I Proteins of p-p65, p65, p-c-jun, and c-jun in cell lysate were detected by western blotting assay. Statistical analysis of interested protein levels was shown. J Proteins of p65, c-jun, and GSDME-FL were detected by western blotting assay in lysate of cell plasma or nucleus, respectively. Statistical analysis of interested protein levels was shown. GAPDH and HDAC1 served as a loading control of cell plasma protein and nucleus proteins, respectively. K, L Immunofluorescence assay was used to detect translocation of p65 and c-jun into nucleus. M Co-immunoprecipitation assay was used to detect the interaction between GSDME-FL and p65 or c-jun. Scale bar represents 100 μm in C, D, H, K and L. ns not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

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