Fig. 5

Effects of I3C on mRNA and protein levels in light-challenged BALB/cJ mice. a Schematic overview of experimental design. Eight to ten-week-old BALB/cJ mice of both sexes were used. Mice were dark-adapted for 16 h before the mice were exposed to white light with an intensity 15,000 lux for 1 h. The animals received intraperitoneal injections of 15 mg/kg I3C or vehicle (DMSO), 1 day before the light exposure and once daily for the remaining 3 days. Four days after the light exposure, b-f mRNA expression levels of i-NOS b, IL-1ß c, NLRP3 d, IL-6 e, and CCL2 f, were analyzed by real-time PCR. Four days after the light exposure, retinal lysates were extracted to perform ELISA of CCL2 g and Western blots h detecting i-NOS, IL- 1ß, p-NFKB65, NQO1, and HMOX1 protein levels. Beta-actin served as loading control. Relative protein levels for i-NOS i, p-NFkBp65 j, and NQO1 k were determined by densitometry. Data show mean ± SEM out of three independent experiments for mRNA expression levels (untreated n = 8 retinas, vehicle + light n = 14 retinas, light + I3C treatment n = 14 retinas, where each dot represents one retina). For Western blots and ELISA three independent experiments were performed (one retina per group) with *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001