Fig. 1

(A-E): Circadian difference in HO-1 protein expression in BV2 cells

(A)  Per-2 mRNA level in microglia cell line BV2 in the late and early phase of the circadian cycle demonstrated as fold change late vs. early phase. Early and late phases in the circadian cycle were induced by different timing of the medium change. (early vs. late phase p = 0.0247, n = 5). (B-D) Western blot showing time-dependent HO-1 protein expression level with or without carbon monoxide (CO) and blood exposure in the microglia cell line BV2. Shown late phase on the left, early phase on the right. Early and late phases in the circadian cycle were induced by different timing of medium change, treatment with 250 ppm CO and/or 100 µl blood for 1 h (h). (B) Representative western blot. (C) Western blot quantification of HO-1 protein expression level early vs. late phase in BV2 cells, p = 0.0197, n = 5. (D) Western blot quantification of HO-1 protein expression level in BV2 cells treated with 100 µl blood for 1 h.early vs. late phase, p = 0.9822, n = 5. (E) Western blot quantification of HO-1 protein expression level in BV2 cells treated with 100 µl blood and 250 ppm CO for 1 h early vs. late phase, p = 0.0128, n = 5, Used statistical analysis: paired t-test; results were presented as box blot with median, minimum and maximum or bars of means with SD. Statistically significant values were defined as p ≤ 0.05 (* p ≤ 0.05; ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001)