Fig. 2: TPSN-sfGFP-3xHA is functional and associates both with PLC and RNF185/MBRL ERAD complex. | Nature Communications

Fig. 2: TPSN-sfGFP-3xHA is functional and associates both with PLC and RNF185/MBRL ERAD complex.

From: Tapasin assembly surveillance by the RNF185/Membralin ubiquitin ligase complex regulates MHC-I surface expression

Fig. 2

a TPSN-sfGFP-3xHA is functional. Cell surface expression of MHC-I was analysed with the conformation-sensitive W6/32 antibody in Parental, TPSN KO, and TPSN KO HEK cells stably expressing TPSN-sfGFP-3xHA using flow cytometry. b Proteins co-precipitating with TPSN-sfGFP-3xHA were analysed by mass spectrometry. The x-axis shows enrichment of proteins associated with TPSN-sfGFP-3xHA over an untagged control. The −log10 of the p-value is shown on the y-axis. p-values were calculated in Perseus using a two-sample t-test for significance. Proteins significantly enriched (FDR < 0.05) are displayed in black, orange (PLC components) or green (quality control factors). Three independent replicates were analysed. c Validation of the proteins co-precipitating with TPSN-sfGFP-3xHA from (b) as analysed by immunoblotting. Cells were lysed in buffer containing 1% DMNG and TPSN-sfGFP-3xHA was immunoprecipitated. Selected proteins were analysed by immunoblotting with the antibodies indicated.

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