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. 2015 Aug 17;112(35):11108–11113. doi: 10.1073/pnas.1415260112

Fig. S2.

Fig. S2.

(A) The abundance of phyB-GFP does not change significantly in seedlings grown under 12h L/12h D cycles. Transgenic phyB-9 seedlings expressing the 35S:PHYB-GFP transgene were grown in 12 h WL or RL/12 h D cycles for 5 d. Samples were harvested at MOD and MON of day 6. Total protein extracts were separated by SDS/PAGE. phyB-GFP and ACTIN proteins were detected using anti-GFP and anti-ACTIN antibodies, respectively. (B) Truncated phyB-YFP fusion proteins lacking Lys996 are not SUMOylated. Seedlings expressing full-length phyB (FL-phyB) and truncated phyB(1–651) or phyB(1–991) proteins fused to GFP or YFP were grown in darkness for 4 d (D) and transferred to WL for 24 h (W24). Fusion proteins were immunoprecipitated using GFP-Trap agarose beads (IP:αGFP). Samples containing comparable amounts of the different fusion proteins were separated by SDS/PAGE. Replicate Western blots were probed with anti-GFP (IB:αGFP) or anti-SUMO1 (IB:αSUMO1) antibodies. (C) Abundance of phyB-GFP and phyBLys996Arg-YFP fusion proteins expressed under the control of LIP1 and 35S promoters is identical pair-wise. Transgenic phyB-9 seedlings expressing the phyB-GFP fusion proteins were grown in darkness for 5 d. Total protein extracts were separated by SDS/PAGE. phyB-GFP and ACTIN proteins were detected using anti-GFP and anti-ACTIN antibodies, respectively. Results were obtained from independent phyBLys996Arg-YFP–expressing transgenic lines, 1 and 2 (Set 1 and Set 2). Separately framed images of the upper panels were produced from different blots.