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. 2022 Mar 9;34(6):2242–2265. doi: 10.1093/plcell/koac083

Figure 8.

Figure 8

BLI associates with WRI1 to repress CLF and recruit SWI3B during regulating WRI1 target genes. A, WRI1 and CLF competitively interact with BLI, as determined in an in vivo Co-IP. Total protein extracts from transformed WT protoplasts carrying both 35S:BLI-mCherry and 35S:CLF-FLAG without or with different amounts of 35S:MYC-WRI1 were immunoprecipitated with the immobilized anti-mCherry antibody. B, WRI1 and SWI3B synergistically interact with BLI, as determined in an in vivo Co-IP assay. Total protein extracts from transformed WT protoplasts carrying both 35S: YFP-WRI1 and 35S:MYC-SWI3B without or with different amounts of 35S:BLI-mCherry were immunoprecipitated with the immobilized anti-GFP antibody. C and D, CLF and SWI3B competitively interact with the BLI–WRI1 complex, as determined in an in vivo Co-IP assay. Total protein extracts from transformed 35S:YFP-WRI1 (C) or 35S:BLI-mCherry (D) protoplasts carrying both 35S:MYC-SWI3B and 35S:CLF-FLAG with different amounts of 35S:BLI-mCherry (C) or 35S:YFP-WRI1 (D) were immunoprecipitated with the immobilized anti-GFP (C) or anti-mCherry (D) antibody. The variable amounts of tagged BLI and WRI1 protein were achieved by adding 1, 2, 3, or 4 times the amount of expression vector, and the proteins from crude lysates (Input) and IPs were detected using different antibodies (A–D). E–I, Occupancy of CLF on the promoters of fatty acid biosynthesis genes BCCP2 (E), KAS1 (F), Cy-PK (G), FAD2 (H), and SUS2 (I) in CLF-GFP, CLF-GFP/bli-1, and CLF-GFP/MYC-BLI seeds compared to GFP control seeds. J–N, Occupancy of SWI3B on the promoters of fatty acid biosynthesis genes BCCP2 (J), KAS1 (K), Cy-PK (L), FAD2 (M), and SUS2 (N) in SWI3B-MYC, SWI3B-MYC/bli-1, and SWI3B-MYC/BLI-GFP seed compared to MYC control seeds. F1–F4 indicate different DNA fragments, as shown in Figure 6B; the TA3 locus was used as a negative control. Data shown are means ±  sd (N = 3). Means of enrichment folds do not significantly differ when they are labeled with the same letter, as determined by Tukey’s HSD test (P < 0.05).