Fig. 3

Confirmation and titration assay using polyclonal α-TOR antibody. a Immunoblot analysis to confirm the specificity of whole serum and purified polyclonal α-TOR antibody. Three-day-old Arabidopsis wild-type and tor-est seedlings were grown in the absence or presence of estradiol for another 5 days. The proteins were separated in 6% SDS-PAGE and immunoblot analysis was conducted using a 1:500 dilution. b Titration of purified polyclonal α-TOR antibody. The antibody was titrated with 1:500 and 1:1000 dilutions using the same plant extracts shown in a, and immunoblot analysis was conducted. c Subcellular localization of TOR proteins. Cytosolic and nuclear fractions were separated and conducted for immunoblot analysis using α-TOR (1:500), α-HSP90 (1:20,000) as a cytosolic marker, and α-histone H3 (1:3000) as a nuclear marker. Molecular size markers (kDa) are represented on the left. Est, estradiol; IB, immunoblot; CBB, Coomassie brilliant blue-stained blot