Fig 1: Trimers, long-OG mix (DP > 8) and flg22 induce MAPK3and MAPK6 phosphorylation. Seedlings were treated for 10 min with trimers (200 µM & 1 mM), long-OG mix (200 µM & 1 mM), flg22 (1 µM), or mock solution (1/2 MS). Each biological replicate is a pool of three wells of seedlings, each containing approximately ten plants. Thirty micrograms of each protein sample was loaded and phosphorylation levels were assessed by immunoblotting using a phospho-p44/42 specific antibody (top). MPK3, MPK4 and MPK6 total protein amounts were assessed using specific antibodies (bottom). The experiment was repeated with a second biological replicate, yielding identical results
Fig 2: AP2C1 and MKP1 control wound-induced MAPK activities. Analysis of wound-induced MPK6, MPK4, and MPK3 kinase activities and protein amounts of leaves from 6-week-old WT, ap2c1, mkp1, and ap2c1 mkp1 plants grown under short-day conditions. (A) MAPK activities were determined after immunoprecipitation by phosphorylation of MBP detected by autoradiography. The entire kinase assay was based on one common master mix containing MBP and ?-ATP. Loading is demonstrated by Coomassie blue staining (CBS); representative lanes are shown. The experiment was repeated twice with similar results. (B) MAPK protein amounts before and after wounding demonstrated by immunoblotting of MPK3, MPK4, and MPK6 from total protein extract using specific antibodies. Loading is demonstrated by Ponceau S staining (Rubisco protein). mpw, Minutes post wounding.
Supplier Page from MilliporeSigma for Anti-AtMPK4 antibody produced in rabbit