Figure 2: T_H1 effector activation in glomerulonephritis requires AKAP9.

(a) Frequency of IFNγ-positive cells, following in vitro PMA/ionomycin stimulation, in CD4⁺ T cell isolates from the spleen and immunizing lymph node of AKAP9^cko/CD4 and AKAP9^wt mice 10 days following induction of anti-GBM nephritis. Data presented as mean frequency of cells ±s.e.m., n=7. (b) As in a for the kidney and draining lymph node. (c) T_H1 cells from AKAP9^cko/CD4 and AKAP9^wt mice were labelled and co-transferred into wild-type animals on day 10 following induction of anti-GBM nephritis. Accumulation was analysed in the kidney 3 h after adoptive transfer. Data are mean percentage of recovered cells ±s.e.m., n=5. (d) As in a for CD4⁺ T cells recovered from the kidney but without PMA/ionomycin treatment before intracellular cytokine staining. Plots are representative FACS panels of IFNγ expression in AKAP9^cko/CD4 and AKAP9^wt control cells. (e) Ox40 expression of kidney CD4⁺ T cells of a, presented as fold isotype staining ±s.e.m. Histogram is a representative of Ox40 staining (x axis log scale). (f) Neutrophil (CD45⁺Ly6G⁺Ly6B.2⁺) and (g) Macrophage (CD45⁺CD11b⁺F4/80⁺) infiltration in kidneys of a, presented as number of cells ±s.e.m. (h) Mean albuminuria at day 0 and 10 after induction of anti-GBM nephritis of (a), presented as mg urinary albumin per g of urinary creatinine ±s.e.m. (i) Left panel: Representative micrographs of PAS staining of kidney sections. Asterisk indicates crescent formation, ‘c’ indicates tubular casts and ‘>’ indicates glomerular thrombosis. Scale bar, 100 μm. Right panel: Median histological scoring based on hypercellularity, glomerular thrombosis, crescent formation, tubular casts and morphologic glomerular irregularity±interquartile range. #P<0.05. NS, not significant.