Figure 3.

CD138⁺ Mϕ in pristane-treated mice are M1-like. B6 mice were injected i.p. with pristane or mineral oil (MO). Peritoneal exudate cells were collected at d14. (A) PEC were stained with antibodies against CD11b, CD138, CD273, CD274, Ly6C, and CD86. CD11b⁺CD138⁺ cells were gated to analyze staining of the other markers.***P < 0.001 by unpaired Student’s t-test (panels 1 and 5) or Welch’s t-test (panels 2, 3, and 4). (B) CD11b⁺CD138⁺ cells were flow sorted, and expression levels of Il10, Chil3, Tnfa, Hif1a, and Pfkl were determined relative to 18 S (Q-PCR). *P < 0.05 by Student’s t-test (panels 2 and 5) or Welch’s t-test (panels 1, 3, and 4). (C) Peritoneal CD11b⁺Ly6C^hi and CD11b⁺CD138⁺ cells were flow sorted from pristane- and MO-treated mice. OCR was measured (XF96 Analyzer). Left, CD138⁺ Mϕ from pristane- vs. MO-treated mice; middle and right, Ly6C^hi vs. CD138⁺ Mϕ from individual pristane- and MO-treated mice. Experimental treatments were performed with six technical replicates. **P < 0.01, ***P < 0.001 by Welch’s unpaired t-test (left) or Student’s paired t-test (middle and right). (D) BODIPY-labeled LDL (10 µg/ml) was added to PEC from pristane- and MO-treated mice for 2 h and cells were then stained with anti-CD11b, Ly6C, and CD138. Mean fluorescence intensity (MFI) of BODIPY-LDL was analyzed. Comparison of Ly6C^hi vs. CD138⁺ Mϕ from individual pristane- (left) and MO- (right) treated mice **P < 0.01; ****P < 0.0001 vs. control, paired Student’s t-test.