Figure 10. HHcy+T2DM induced Ly6C expression possibly via CEBPα induction.

(A) Schematic illustration of transcriptome screening. (B) Sorting strategy for CD11b⁺Ly6G^-Ly6C^high inflammatory MCs (Ly6C^high) and CD11b⁺Ly6G^-Ly6C^low resident MCs (Ly6C^low) in WT mouse blood. Each population obtained 200,000 cells and RNA was extracted for RNA sequencing. (C) Upregulated TFs in Ly6C^high cells. Using HiSeq X Ten system, we obtained 30 million reads from each sample, among which 17,230 genes were recognized. Ly6C^high population had 1,776 genes upregulated compared with Ly6C^low group. Nine upregulated transcription factors were identified within Ly6C^high 1,776 gene list. (D) Ly6c promoter analysis. Two core promoter regions (−1348/−947, −348/454, blue rectangle) were predicted by Encode. CEBPα transcription factor binding sites (−957/−952, −236/−231, grey diamond) were identified in mouse Ly6c core promoter regions by PROMO. (E) Frequency of CEBPα⁺ cells among total Ly6C⁺ MCs. N=4 mice. *P<0.05 vs CT diet mice with the same genotype. Synergy was defined as HHcy and T2DM produced a greater effects in db/db mice on HM diet than the sum of that in HHcy and T2DM alone. WBC, white blood cell.