Fig. 1: The expression of RNF128 in macrophages is augmented during atherogenesis.

A Subset of cells from atherosclerotic aortas was annotated into nine clusters. B Location of cell clusters with Rnf128 expression on the t-SNE plot. C Cell clusters with the expression of Rnf128 (Rnf128⁺Mφ, left), high expression of Lyz2 (Lyz2^hiMφ, middle), and both of these two genes (Rnf128⁺Lyz2^hiMφ, right). Mφ, macrophages. D The expression level of Rnf128-expressing macrophage numbers during the process of atherosclerosis. E The number of macrophages with high expression of Rnf128 and Lyz2 in atherosclerotic aortas of mice fed WD for different durations. F Colocalization analysis via immunofluorescence of RNF128 and MOMA-2 (specific for monocytes and macrophages) expression in early and advanced atherosclerotic lesions of apolipoprotein E null (ApoE^−/−) mice fed a WD for 8 weeks and 20 weeks, respectively (n = 8 per group, hereafter n = 8). Scale bar: 100 µm. G Western blotting images of RNF128 protein levels from whole aortas and quantitative analysis (n = 6). H Colocalization analysis of RNF128 and MOMA-2 expression in early and advanced atherosclerotic lesions from coronary atheromatous plaques of humans (n = 8). Scale bar: 100 µm. I Immunofluorescence of RNF128 in macrophages incubated with oxidized low-density lipoprotein (oxLDL, 75 µg/mL) for different time points (n = 6). Scale bar: 20 µm. J Western blotting (left, n = 4) and quantitative PCR (right, n = 6) of RNF128 expression in macrophages treated with oxLDL for different time points. K Western blotting (left, n = 4) and quantitative PCR (right, n = 6) of RNF128 expression in macrophages treated with a concentration gradient of oxLDL for 24 h. L, M Western blotting (left, n = 4) and quantitative PCR analysis (right, n = 6) of RNF128 expression in RAW264.7 and THP-1-derived macrophages treated with time-dependent oxLDL, respectively. The “n” represents the number of biologically independent samples. Data were presented as mean ± SD, Shapiro–Wilk method tested that all data were normally distributed. Unpaired two-tailed Student’s t-test was used for (G). One-way ANOVA followed by the Dunnett post hoc test was used for the others. Adjusted P values were provided in case of multiple-group comparisons. Source data are provided as a Source Data file.