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Fig. 7 | BMC Biology

Fig. 7

From: SKIP controls flowering time via the alternative splicing of SEF pre-mRNA in Arabidopsis

Fig. 7

SKIP is required for the pre-mRNA splicing of SEF through direct binding, and it regulates the deposition of H2A.Z at FLC, MAF4, and MAF5 chromatin. a Phenotypes of the plant materials used for RNA-IP. b RNA-IP analysis of WT and C12-1 plants. SKIP-associated SEF pre-mRNA was detected by RT-PCR. C12-1 is a skip-1 complemented transgenic line harboring the SKIP:GFP-SKIP construct. ACT2 was used as a negative control. Input, positive control; NA-IP, no antibody was used during immunoprecipitation as a negative control; GFP-IP (RT+) or GFP-IP (RT–), PCR was performed following RNA-IP against GFP with (RT+) or without (RT–) reverse transcription. c Global changes in H2A.Z deposition in WT, skip-1, sef-2, and arp6-5 plants as detected by Western blot using anti-H2A.Z antibodies. Tubulin was used as a loading control. The relative level of H2A.Z to tubulin was quantified by Image J software. One-way analysis of variance (ANOVA; Tukey’s multiple comparison test) was performed. Statistically significant differences are indicated by different lowercase letters (P < 0.05). d, f H2A.Z enrichment at FLC (d), MAF4, and MAF5 (f) chromatin in WT, skip-1, sef-2, and arp6-5 plants as measured by ChIP with qPCR using anti-H2A.Z antibodies. The data are the mean ± s.d. (n = 3) in d and f. e, g Diagram of FLC (e), MAF4, and MAF5 (g) with exons indicated as black boxes, introns indicated as gray boxes, and the promoter indicated as a black line. The PCR primer sets used are shown as black lines below the diagram. The names of the primer sets correspond to the numbers on the x-axis of the graphs in d and f. kb kilobases

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