Figure 6.

EC2319’s anti-monocyte activity in anti-GBM rats and EAU mice

(A) Glomerulonephritis was induced in Wistar Kyoto (WKY) rats (n = 3) on day 0 with anti-GBM antibody, followed by EC2319 treatment (8 h and on day 3) and fluorescence-activated cell sorting (FACS) analyses, as indicated.

(B) CD172⁺ CD43^- monocytes found in PBMCs isolated from anti-GBM rats on days 6 and 12 post induction and compared to healthy littermates.

(C) CD172⁺ CD43⁻ monocytes as percentage of total blood leukocytes isolated from anti-GBM rats treated with EC2319 (575 nmol/kg) without or with a 300-fold molar excess of the FA competitor or oral MTX in two ways.

(D) Flowchart illustrating a daily dosing schedule of EC2319 (subcutaneous) and BLZ945 (oral gavage) in EAU mice (n = 10).

(E) Flow cytometry gating strategy on CD45⁺ CD11b⁺ NK1.1⁻ mouse leukocytes for identification of classical (R1, Ly6C^high CD43⁺) and intermediate (R2, Ly6C^high CD43^high) monocytes.

(F) Mean effect of EC2319 and BLZ945 treatment on R1 and R2 monocyte subpopulations per 100 μL mouse blood as bar graphs with healthy controls (no EAU) and individual data points.

Values represent mean ± SEM. One-way ANOVA with Tukey’s post hoc analysis: ∗∗∗p < 0.001 and ∗p < 0.05 for the comparing groups in (B) and (C). Kruskal-Wallis test with Dunn’s post hoc analysis: ∗∗∗p < 0.001 and ∗∗p < 0.01 in (F). ns, not significant.