Fig. 1: Genetic deletion or pharmacological inhibition of MIF−2 attenuates early atherosclerotic lesions and vascular inflammation in vivo.

a Experimental outline: Female Apoe^–/– and Mif-2^–/–Apoe^–/– mice fed a cholesterol-rich HFD for 4.5 weeks. b ORO staining of aortic root sections and corresponding quantification (12 serial sections per mouse; n = 12 mice; each data-point represents one mouse; scale bar: 250 µm). c Same as (b) but HE staining. d HE staining and quantification of aortic arch sections (8 sections per mouse; n = 12 mice; scale bar: 200 µm). e Same as (b, c), except that sections were stained for CD68⁺ macrophages (red) and DAPI (blue); scale bar: 200 µm. f–h Quantification of 40 inflammatory/atherogenic cytokines/chemokines in plasma samples by mouse cytokine array (n = 4 mice per group, analyzed in duplicate each). f Heatmap illustrating altered levels of cytokines/chemokines. Signals from Apoe^–/– were normalized to 1 and compared with those from Mif-2^–/–Apoe^–/– mice (upregulated cytokines in magenta, downregulated in cyan). g, h Bar graphs of significantly altered cytokines/chemokines with relatively lower (h) or higher (g) plasma abundance. i–m Inhibition of MIF-2 by 4-CPPC in a 4.5-week HFD male Apoe^–/– model. i Scheme of the experimental outline. Mice were administered 4-CPPC (5 mg/kg) or vehicle 3× per week (Syringe icon was created with BioRender.com). j ORO staining of aortic root sections and quantification (12 sections per mouse; n = 11 mice per group; scale bar: 200 µm). k Same as (j) but HE staining. l Same as (j), except that sections were stained for CD68⁺ macrophages (red) and DAPI (blue). m Quantitation of plasma cytokines/chemokines. Heatmap illustration of the 36 ProcartaPlex mouse cytokine/chemokine array (n = 4 mice per group analyzed in duplicate). Signals from vehicle-treated mice normalized to 1 and compared with those from 4-CPPC-treated mice (upregulated cytokines in magenta, downregulated in cyan). a and i were created in BioRender. Bernhagen, L. 2025 https://BioRender.com/c59v181. Values are expressed as means ± SD and statistically analyzed using an unpaired two-tailed t-test (b–e and j–l) or multiple unpaired t-tests (g, h).