Extended Data Fig. 7: ABCA1 mediates the functional impact of WDR4 on cholesterol efflux and tumour-promoting activity in TAMs.
a, Total cholesterol levels in WT and Abca1-KO iBMDM-derived TAMs with or without Wdr4 KO (n = 3 biological replicates), and in THP-1-derived TAMs with knockout of ABCG1, APOA1, APOA2, or APOA4, with or without Wdr4 KO (n = 3). b, Membrane cholesterol levels in TAMs derived from THP-1 cells or iBMDMs as detected by cholera toxin B (CTB) staining (n = 3 biological replicates). CTCF, corrected total cell fluorescence. c, NBD-cholesterol efflux percentage in TAMs derived from THP-1 cells or iBMDMs (n = 3 biological replicates), quantified by fluorescence intensity in cells and medium. d, Immunoblotting analysis of ABCA1 expression in ABCA1-overexpression (OE) TAMs (n = 3 biological replicates). e,f, Flow cytometry analysis of iNOS (e) and Arg1 (f) expression in WDR4-KO TAMs transfected with vector or ABCA1-OE plasmid (n = 3 biological replicates). g, Schematic diagram of the co-culture of Huh-7 cells with THP-1-derived TAMs. Created with BioRender.com. h–j, CCK8 assay (h), colony formation assay (i), and EdU assay (j) were performed to assess whether ABCA1 OE could rescue the WDR4-KO TAM-induced inhibition of Huh-7 cell proliferation (n = 3 biological replicates). k, Tumour growth curves, and final tumour weights in WT C57BL/6 mice subcutaneously co-injected with Hepa1-6 cells and Wdr4-KO iBMDMs transfected with ABCA1-OE or vector (n = 5 mice per group). All values are presented as mean ± s.d. Statistical significance was calculated using two-tailed unpaired Student’s t-test (quantifications in d), two-way ANOVA (a–c,h, tumour growth curves in k), or one-way ANOVA (i,j; quantifications in e,f,k) with Tukey’s post-hoc test.
