Densities of MAP1LC3-II were normalized to control treatment and relative fold changes of MAP1LC3-II were normalized to GAPDH. ( C ) Immunoblot analysis of MSCs starved for 0 (control) to 12 h, or treated with TNF (20 ng/ml) or IFNG (50 ng/ml) for the indicated times. Sections were stained with DAPI and examined by confocal microscopy. Mice were euthanized at 24, 48, 72, and 120 h later, the lungs and spinal cords were separated and cut into frozen sections. ( A and B ) MAP1LC3-EGFP-MSCs were injected into naive and EAE (on day 15) mice via tail vein. MSCs undergo autophagy in the inflammatory microenvironment. : not significant, compared to WT, t -test).įigure 1. Data presented are the mean +- SEM of the relative cell numbers compared to MRL-Fas lpr KSRP +/+ mice (19 to 23 animals per genotype) (**: p < 0.01, n.s. ( c - e ) FACS was used to analyze intra-renal frequency of CD4 +, CD4 + -IFN-gamma + and FoxP3 + cells in the kidneys of female 15-week-old MRL-Fas lpr KSRP +/+ and MRL-Fas pr KSRP -/- mice. Representative image of immunostaining with 40x magnification. Data presented are the mean +- SEM of the score with 10 animals per genotype (**: p < 0.01, *: p < 0.05 compared to WT, TTEST). ( b ) Immunostaining was used to analyze intra-renal frequency of FoxP3, Ki67 and IFN-gamma producing cells in the kidneys of female 19-week-old MRL-Fas pr KSRP +/+ and MRL-Fas lpr KSRP -/- mice. : not significantly different from WT, T -TEST). Data presented are the mean +- SEM of the relative mRNA expressions based on WT female 19 weeks (100%) 9 to 14 animals per genotype (*: p < 0.05 n.s. T cell lineage-specific transcription factor mRNA expression of T-bet, GATA3, RORgammaT and FoxP3 were analyzed by qRT-PCR and normalized to the mRNA expression of Pol2a. Figure 4 Characterization of T cell population in the kidneys of 19 and 15-week-old female MRL-Fas lpr KSRP -/- mice compared to MRL-Fas lpr KSRP +/+ mice ( a ) RNA was isolated from the kidneys of 19-week-old female MRL-Fas lpr KSRP -/- mice (KO) and MRL-Fas lpr KSRP +/+ mice (WT).
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