Fig. 1.

GSK4112 inhibited LPS-induced BV2 cell activation. a HEK293 cells were transfected with Bmal1 and Renilla luciferase reporter and then treated with DMSO or GSK4112 (40 μM) for 16 h. The value of the group without drug (dissolvent only) was set to 1. *P < 0.05, n = 3. b BV2 cells were pretreated with DMSO or GSK4112 (40 μM) for 16 h. Cell lysates were collected to measure Bmal1 mRNA levels by qRT-PCR assay. *P < 0.05, n = 3. c Normalized BV2 cell viability influenced by different dose of GSK4112 (0–80 μM) for 16 h as assessed by MTT assay. d BV2 cells were pretreated with different doses of GSK4112 (2.5–40 μM) for 4 h and then exposed to LPS (100 ng/mL) for 12 h. Cell lysates were analyzed by Western blot for iNOS, COX-2, and GAPDH. The relative band intensity of iNOS and COX-2 to GAPDH were analyzed. *P < 0.05, **P < 0.01, ***P < 0.001, n = 3. e BV2 cells were pretreated with different doses of SR9011 (0–10 μM) for 4 h and then exposed to LPS (100 ng/mL) for 12 h. Cell lysates were analyzed by Western blot for iNOS, COX-2, and GAPDH. The relative band intensities of iNOS and COX-2 to GAPDH were analyzed. **P < 0.01, ***P < 0.001, n = 3. f, g BV2 cells were pretreated with DMSO or GSK4112 (40 μM) for 4 h and then exposed to LPS (100 ng/mL) for 12 h. Cell lysates were collected to measure iNOS, COX-2, IL-6, and TNFα mRNA levels by qRT-PCR assays. Cultured media were collected to measure IL-6 and TNFα levels by ELISA assays. *P < 0.05, **P < 0.01, ***P < 0.001, n = 3