Fig. 2

Inhibition of CD147 attenuates early proinflammatory activation of Ly-6C+ monocytes/macrophages (MMs) in the spleen at 4 h tMCAO. a Representative western blot images showing protein levels of CCR2 and Ly-6C detected in isolated splenocytes from the following groups (n = 5 mice/group): sham, tMCAO, tMCAO + isotype, and tMCAO + αCD147. Similar results were obtained in three independent experiments. No difference was noted between tMCAO and +iso groups. b RT-qPCR analysis of mRNA levels of CCR2 and Ly-6C detected in isolated splenocytes from the following groups (n = 5 mice/group): sham surgery, tMCAO + isotype, and tMCAO+αCD147. *p < 0.05 vs. sham control; ^#p < 0.05 vs. tMCAO + isotype. c Representative dot plots illustrating the flow cytometry gating strategy for identification of immune cell subsets in the spleen. Monocyte subsets in the spleen were identified by selecting the gate (R1) APC-CD11b^high and PE-Lin^low and further separated by CD11c/F4/80-PeCy5 and Ly-6C-FITC. d Flow cytometric quantification of Ly-6C^high and Ly-6C^low monocyte subsets. The number and ratio of Ly-6C^high and Ly-6C^low monocyte subsets in spleen were measured in the following groups (n = 5 mice/group): sham surgery, tMCAO + isotype, and tMCAO + αCD147. *p < 0.05 vs. sham; ^#p < 0.05 vs. tMCAO + isotype. e RT-qPCR analysis of mRNA levels of iNOS, IL-6, and Arg1 detected in the FACS-sorted splenic monocytes from the indicated groups of mice (n = 6/groups). *p < 0.05 vs. sham control; ^#p < 0.05 vs. tMCAO + isotype