Fig. 2

BAF60 expression is regulated by both light and circadian rhythm. a RT-qPCR of Baf60 expression in a 7-day-old wild-type line (Ws) grown in LD (7-d-old LD) or in darkness (7-d-old darkness) conditions. Values are average ± standard deviation obtained from three independent replicates. Asterisks indicate significantly different values (Student’s t-test, P < 0.05). b RT-qPCR data showing the relative expression of Baf60 in wild-type plants under LD conditions in comparison to TOC1, a circadian clock-regulated gene. Total RNA samples were collected every 4 h from 14-day-old plants during a 32-h period (Light and Dark represent day and night periods, respectively). The grey area behind the trace represents the night period. Values are average ± standard deviation obtained from three independent replicates. c Immunoblot analysis showing the amount of BAF60 protein under light and dark conditions, obtained from a promBAF60::BAF60-CFP line. d Immunoblot analysis showing that MG132 can prevent BAF60 degradation through proteasome 26S in the same ratio under light and dark conditions. This result indicates that BAF60 degradation is independent of light. e RT-qPCR data showing the relative expression of BAF60 in wild-type plants under continuous light. The seedlings were grown under LD conditions and were then transferred to continuous day conditions for the indicated period of time. Total RNA samples were collected every 4 h from 14-day-old plants during a 20-h period. The grey area behind the trace represents the period that would have corresponded to the night. Values are average ± standard deviation obtained from three independent replicates. f RT-qPCR quantification of BAF60 expression in 7-day-old wild type seedlings grown in LD conditions and then transferred to darkness (3hrs Dark, grey) or kept in the light (Light, white) for 3 h. Values are average ± standard deviation and are representative of three biological replicates. Asterisks indicate significantly different values (Student’s t-test, P < 0.05). g BAF60 promoter activity observed by RT-qPCR quantification of GFP expression in 7-day-old BAF60::WPP-GFP-BLRP seedlings grown in LD conditions and then transferred to darkness (3hrs Dark, grey) or not (Light, white) for 3 h. Values are average ± standard deviation obtained from three independent replicates. Asterisks indicate significantly different values (Student’s t-test, P < 0.05)