Figure 6
Protein–protein interaction between PmlR2 and PmSB-LOV by gel-filtration chromatography. The recombinant PmlR2 (A), PmSB-LOV (B), PmSB-LOVC53A and PmlR2 (C), and PmSB-LOV and PmlR2 (D) incubated under dark (solid line) and light (dashed line) conditions were applied to a Superdex 200 HR 10/30 column on an ÄKTA FPLC system. PmlR2 (200 µL; 0.43 mg/mL), PmSB-LOV (200 µL; 0.47 mg/mL), PmSB-LOVC53A (200 µL; 0.21 mg/mL) recombinant proteins were used. The column was equilibrated and developed with 1 × PBS at a flow rate of 0.25 mL/min. Molecular size standards (Fe: Ferritin, Al: Conalbumin, and Ca: Carbonic anhydrase indicating 440, 75, and 29 kDa, respectively) were applied. White light was applied at approximately 50 µmol s−1 m−2 for 5 min. SDS-PAGE and silver staining for panel D is shown at the bottom. Two independent experiments in technical replicates were performed, and representative data were shown. The original and unprocessed versions of the full-length stained gels were included in Fig. S13.
