S where phenotyped on MS medium containing 0 and 3 (w/v) Suc and soil (peat/perlite/vermiculite) according to the parameters described previously (Boyes et al., 2001). Seeds were sterilized with chlorine gas and sown on MS medium containing 3 (w/v) Suc followed by 48 h of stratification at 4 . Plants were grown for 2 weeks at 22 with a light intensity of 80 mmol quanta m22 s21 in a 16-h photop
S where phenotyped on MS medium containing 0 and 3 (w/v) Suc and soil (peat/perlite/vermiculite) according to the parameters described previously (Boyes et al., 2001). Seeds were sterilized with chlorine gas and sown on MS medium containing 3 (w/v) Suc followed by 48 h of stratification at 4 . Plants were grown for 2 weeks at 22 with a light intensity of 80 mmol quanta m22 s21 in a 16-h photop
Es and one proline of the CxxxPxC motif located upstream of the CxCxxC clusters did reduce PP2A binding significantly. Additional studies that relied upon peptide inhibition of the tAg-PP2A interaction without destabilizing tAg suggested that the CxCxxC clusters might indeed influence tAg binding to the A subunit [23,25]. Two recent X-ray crystallographic studies indicate the CxCxxC clusters and t
Es and one proline of the CxxxPxC motif located upstream of the CxCxxC clusters did reduce PP2A binding significantly. Additional studies that relied upon peptide inhibition of the tAg-PP2A interaction without destabilizing tAg suggested that the CxCxxC clusters might indeed influence tAg binding to the A subunit [23,25]. Two recent X-ray crystallographic studies indicate the CxCxxC clusters and t
Mitochondrial pellet was resuspended in digitonin buffer (30 mM HEPES-KOH, pH 7.4, 150 mM potassium acetate, 10 [v/v] glycerol, and 1 mg of digitonin) and kept on ice for 30 min. Ten microliters of antibody serum, 50 mL of Protein A-Sepharose, 1 (w/v) BSA, and Complete protease inhibitor (Roche) were added to the mitochondria and incubated for 6 h at 4 on a rotating wheel at 20 rpm. Beads were
Mitochondrial pellet was resuspended in digitonin buffer (30 mM HEPES-KOH, pH 7.4, 150 mM potassium acetate, 10 [v/v] glycerol, and 1 mg of digitonin) and kept on ice for 30 min. Ten microliters of antibody serum, 50 mL of Protein A-Sepharose, 1 (w/v) BSA, and Complete protease inhibitor (Roche) were added to the mitochondria and incubated for 6 h at 4 on a rotating wheel at 20 rpm. Beads were
Flanked by Gateway recombination cassettes (Supplemental Table S4) and cloned intoMurcha et al.pDONR201. LR reactions were carried out with C-terminal GFP fusion vectors for GFP localization (Carrie et al., 2009), pDEST14 for in vitro transcription and translation, and pB2GW7 (35S CaMV promoter) for Agrobacterium tumefaciensmediated complementation (Karimi et al., 2002). Yeast two-hybrid vectors w
Mitochondrial pellet was resuspended in digitonin buffer (30 mM HEPES-KOH, pH 7.4, 150 mM potassium acetate, 10 [v/v] glycerol, and 1 mg of digitonin) and kept on ice for 30 min. Ten microliters of antibody serum, 50 mL of Protein A-Sepharose, 1 (w/v) BSA, and Complete protease inhibitor (Roche) were added to the mitochondria and incubated for 6 h at 4 on a rotating wheel at 20 rpm. Beads were