Fig. 6
Mmp12–/– mouse mortality and IFN-γ macrophages numbers in SLE. a Following CFA injection, the size of the superficial cervical lymph nodes of Mmp12+/+ and Mmp12–/– MRL/lpr (n = 16 and 12, respectively, for each time point) female mice were measured and expressed as means ± s.d. Two-tailed unpaired Student’s t-test: ***p < 0.005. b Kaplan–Meier curves showing mortality rates of female Mmp12+/+ MRL/lpr (n = 16) and Mmp12–/– (n = 12) mice. Two-tailed unpaired Student’s t-test: p < 2 × 10−10, (see Supplementary Fig. 9c, d for survival data). c Representative images of superficial cervical lymph nodes (white arrows) immunostained for IFN-γ, iNOS, MHCII, and CD36 from Mmp12+/+ (n = 3) and Mmp12–/– (n = 3) female MRL/lpr mice at their humane end points (Mmp12+/+ (day 112) and Mmp12–/– (day 98)). Scale bars, 100 μm. d Quantification of immunostaining intensities of Mmp12+/+ (n = 3) versus Mmp12–/– (n = 3) MRL/lpr mice superficial cervical lymph nodes for IFN-γ (p < 9 × 10−6), iNOS (p < 2 × 10−7), MHCII (p < 1 × 10−2), and CD36 (p < 2 × 10−2) expressed as the mean ± s.d. Two-tailed unpaired Student’s t-test: *p < 0.05, **p < 0.01, ***p < 0.005. e Superficial cervical lymph nodes were analyzed by western blotting for macrophages markers of IFN-γ activation (iNOS, MHCII, S100A8, and S100A9) and STAT1, as well as for IL-4 induced CD36 and STAT6. Two biological replicates analyses from Mmp12+/+ (n = 8) and Mmp12–/– (n = 8) MRL/lpr mice, are shown. Red arrows indicate lower molecular forms of IFN-γ. f Mean kidney weights and g mean histological scores of activity and chronicity indexes from Mmp12+/+ (n = 11) and Mmp12–/– MRL/lpr (n = 15) mice were measured at their humane end points. Error bars, s.d. Two-tailed unpaired Student’s t-test: *p < 0.05, ***p < 0.005. Quantification of immunostaining in h glomeruli and i in glomeruli and interstitium of Mmp12+/+ (n = 3) and Mmp12–/– MRL/lpr (n = 3) mice at their humane end points for IFN-γ (p < 2 × 10−12), MHCII (p < 7 × 10−17), and CD36 (p < 2 × 10−6). Quantification is expressed as means ± s.d. Two-tailed unpaired Student’s t-test: ***p < 0.005. j Representative images of H&E, IFN-γ, MHCII, and CD36 immunostaining of the renal glomeruli sections of Mmp12+/+ and Mmp12–/– female MRL/lpr mice; arrowheads, strong IFN-γ staining. Scale bars, 100 μm
