Fig. 5: RNF128 interacts with S100A8. | Cell Death & Disease

Fig. 5: RNF128 interacts with S100A8.

From: RNF128 deficiency in macrophages promotes colonic inflammation by suppressing the autophagic degradation of S100A8

Fig. 5

A THP-1 cells were transfected with empty vector or RNF128-Flag. The cell lysates were immunoprecipitated with an anti-FLAG antibody. S100A8 was identified via mass spectrometry. B Mass spectrometry analysis of S100A8 peptide immunoprecipitated by RNF128. C Total THP-1 cell lysates were immunoprecipitated with anti-RNF128 or anti-S100A8 antibodies. RNF128 and S100A8 were detected by western blot. D 239T cells were co-transfected with RNF128-Flag and S100A8-GFP for 48 h. Total cell lysates were immunoprecipitated with anti-Flag or anti-GFP antibodies. RNF128-Flag and S100A8-GFP were detected by western blot. E Immunofluorescence assay of RNF128 and S100A8 in THP-1 cells. Representative confocal microscopy images were shown. Scale bars, 5 μm. F The colocalization of RNF128 and S100A8 in THP-1 cells was analyzed. G Schematic diagram of RNF128-Flag and its truncation mutants. H 239T cells were co-transfected with RNF128-Flag, RNF128-Flag1-276, RNF128-Flag277-428 and S100A8-GFP for 48 h. Total cell lysates were immunoprecipitated with an anti-Flag antibody. The immunoprecipitation complex was detected with anti-GFP and anti-Flag antibodies. I Schematic diagram of S100A8-GFP and its truncation mutants. J 239T cells were co-transfected with S100A8-GFP, S100A8-GFP1-46, S100A8-GFP47-93 and RNF128-Flag for 48 h. Total cell lysates were immunoprecipitated with an anti-GFP antibody. The immunoprecipitation complex was detected by anti-GFP and anti-Flag antibodies.

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