Fig. 2: B7-H4 palmitoylation prevents its lysosomal degradation.
From: Palmitoylation prevents B7-H4 lysosomal degradation sustaining tumor immune evasion
a Normalized heatmap showing screen results of small molecules targeting post-translational modifications in MDA-MB-468 cells. B7-H4-GFP expression was quantified by flow cytometry. NT no target. Immunoblot for B7-H4 in MDA-MB-468 (b), T-47D (c), and 4H11 (d) cells treated with 2-Bromopalmitic acid (2-BP) or palmostain B at the indicated concentrations for 24 h. Immunoblot for B7-H4 in mouse mammary tumor virus-polyoma middle tumor antigen (MMTV-PyMT) cells treated with 2-BP (e) or cerulenin (f) at the indicated concentration for 24 h. Biotin-acyl-exchange (ABE) detects endogenous B7-H4 palmitoylation in MDA-MB-468 (g) and 4H11 (h) cells with or without (w/o) 2-BP (100 μM) treatment for 24 h. B7-H4 palmitoylation levels were measured relative to the corresponding input levels of B7-H4 protein. Representative immunoblot for B7-H4 and protein level quantification from three independent tests in MDA-MB-468 (i) and 4H11 (j) cells examined by cycloheximide (CHX) chase assay combined w/o 2-BP (100 μM) treatment. Data are shown as mean ± SEM. Statistical analyses were performed using two-way ANOVA with Sidak’s multiple comparisons test. Immunoblot for B7-H4 in MDA-MB-468 (k), T-47D (l), and 4H11 (m) cells treated with 2-BP and/or bafilomycin A1 (100 nM) and E64d (20 μM) plus pepstatin A (pep A) (10 μM) for 24 h. Source data are provided as Source Data File.
