Fig. 3: N-Glycosylation of B7H3 stabilizes B7H3 protein in TNBC cells. | Nature Communications

Fig. 3: N-Glycosylation of B7H3 stabilizes B7H3 protein in TNBC cells.

From: FUT8-mediated aberrant N-glycosylation of B7H3 suppresses the immune response in triple-negative breast cancer

Fig. 3

a, b MDA-MB-231 and HCC1806 cells were treated with 20 μM CHX at indicated intervals in the presence of TM (2.5 μg/ml) or not. The intensity of B7H3 protein was quantified using ImageJ software. c The indicated cell lines were treated with 20 μM CHX at indicated intervals. The intensity of B7H3 protein was quantified using ImageJ software. d B7H3-8NQ mutant re-expressing in MDA-MB-231-B7H3KO cells were treated with MG132 (20 μM) in the presence of CHX (20 μM) at indicated intervals. The intensity of B7H3 protein was quantified using ImageJ software. e HEK293T cells were transiently transfected with the indicated plasmids with or without MG132 treatment for 6 h. Immunoprecipitation analysis of exogenous B7H3-8NQ ubiquitination with the indicated antibodies. f B7H3-WT re-expressing in MDA-MB-231-B7H3KO cells were treated with TM for 24 h. Immunoprecipitation analysis of exogenous B7H3 ubiquitination with the indicated antibodies. g Flow cytometry measuring B7H3 protein on the cell membrane with tunicamycin at different concentrations for 24 h. h Flow cytometry measuring B7H3 protein on the cell membrane in the indicated cell lines. The p value in (a–c) was determined by a two-tailed unpaired Student’s t test. Error bars represent mean ± SD. All data are representative of three independent experiments. Red closed circle, glycosylated B7H3; blue star, non-glycosylated B7H3.

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