Figure 4. Effect of anti–miR-93-5p therapy on the lymphoid and myeloid lineages.

(A) Flow cytometric characterization of circulating lymphoid and myeloid cells in 4 different mouse groups: control (no intervention), sham-operated, CLP-induced sepsis with scrambled miRNA treatment, and CLP-induced sepsis with anti–miR-93-5p treatment. Treatment was administrated 24 hours before and 2 hours after sepsis induction. Mice were sacrificed 24 hours after surgery (sham) or CLP-induced sepsis. Control mice were sacrificed together with mice in the other 3 groups. (B) Percentage of CD4⁺CD44⁺CD62L^lo/neg Tem cells (CD4⁺ Tem) in control mice, sham-operated mice, CLP-induced septic mice treated with scrambled miRNA, and CLP-induced septic mice treated with anti–miR-93-5p mice. (C) Percentage of CD8⁺ Tem cells (CD8⁺ Tem) in all 4 groups. (D) Percentage of CSF1R⁺PD-L1⁺ cells in the entire pool of CD11b⁺CSF1R⁺ monocytes in control mice, sham-operated mice, CLP-induced septic mice treated with scrambled miRNA, and CLP-induced septic mice treated with anti–miR-93-5p. (E) Percentage of LyC^hi cells in the entire pool of CD11b⁺CSF1R⁺ monocytes and of (F) LyC^hiPD-L1⁺ monocytes in control mice, sham-operated mice, CLP-induced septic mice treated with scrambled miRNA, and CLP-induced septic mice treated with anti–miR-93-5p. (G) Percentage of F4/80⁺MRC1⁺ macrophages and of (H) F4/80⁺PD-L1⁺ macrophages in all 4 experimental mouse groups. (I) Schematic representation of the effect of anti–miR-93-5p therapy on hematopoiesis during sepsis. Blue rectangles mark the cell subtypes that were significantly differentially expressed in the anti–miR-93-5p–treated group versus the scrambled miRNA–treated group after adjustment for multiple testing using the FDR. Data are presented as the mean ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001, by 2-tailed Student’s t test (B–H). P values that are significant after adjustment for multiple testing using the FDR are highlighted in blue.