Fig. 7: HSD promoted the differentiation and function of M-MDSCs by p38/MAPK-dependent NFAT5 expression.

a Significant Gene Ontology (GO) biological process terms associated with M-MDSCs from HSD-fed mice. Cellular processes enriched by the DEGs in M-MDSCs from 4T1 tumour-bearing HSD- or NSD-fed mice. The p value was tested by DAVID software using a hypergeometic test method. n = 5 mice for the NSD group and 6 mice for the HSD group. b Tumour M-MDSCs were stimulated with an additional 40 mM NaCl for 1 h, and phosphorylated p38 (p-p38) was analysed by western blotting. Right panels: quantification data. c, d Tumour M-MDSCs were transfected with p38-specific siRNA or control siRNA for 2 days, and cultured with an additional 40 mM NaCl for 24 h. Phosphorylated JNK (p-JNK) was analysed by western blotting (right panels: quantification data), and NF-κB activity was determined by the p65 subunit DNA-binding ability. e Two TFs identified as upregulated DEGs in tumour M-MDSCs in 4T1 tumour-bearing HSD-fed mice. f ATF2 and NFAT5 mRNA level in tumour M-MDSCs in 4T1 or B16F10 tumour-bearing HSD- or NSD-fed mice was tested by qRT-PCR. g NFAT5 expression in M-MDSCs from tumour-bearing HSD- or NSD-fed mice was analysed by western blotting. Right panels: quantification data. h An equal number of tumour M-MDSCs were cultured for 24 h in the absence or presence of an additional 40 mM NaCl. NFAT5 expression was determined by western blotting. Right panels: quantification data. i Tumour M-MDSCs were transfected with p38-specific siRNA for 2 days, and cultured in the absence or presence of an additional 40 mM NaCl for 24 h. The level of NFAT5 was examined by western botting. Right panels: quantification data. For b–d and f–h, two-tailed Student’s t test; ns not significant; *p < 0.05, **p < 0.01 and ***p < 0.001; n = 5 mice for the NSD group and 6 mice for the HSD group. Bars are expressed as the means ± SEM of 3 independent replicates with isolated cells pooled from the same group of mice. For i, n = 6 mice. **p < 0.01 and ***p < 0.001. The data are expressed as the mean ± SEM of independent three replicates; one-way ANOVA with post hoc Bonferroni correction. Experiments were repeated three times. Source data are provided as a Source Data file.