Figure 2.

TAP of Arc Reveals Postsynaptic Complexes with a Native Size of 1.5 MDa

(A) BNP of WT and Arc^TAP/+ forebrain extracts blotted with FLAG and Arc antibodies, where Arc and TAP-tagged Arc can be mainly detected at 1.5 MDa. The closed arrow indicates the main Arc complex, whereas open arrows indicate lower-molecular-weight Arc complexes.

(B) Arc was tandem affinity-purified from Arc^TAP/TAP forebrain extracts, eluted, and collected in 5 consecutive fractions following histidines affinity tag (HAT) purification.

(C) Colloidal Coomassie staining of three independent TAPs from WT (left) and Arc^TAP/TAP (right) forebrains. The lanes were cut for LC-MS/MS, and the identified proteins are listed in Table S1. Arc and the Tev enzyme are indicated.

(D) The absolute expression value of each protein in the tandem purification was estimated by the iBAQ intensity values obtained in each purification.

(E) BNP of WT, Arc^TAP/+, and PSD95^TAP/+ forebrain extracts blotted with FLAG antibody. TAP-tagged Arc and TAP-tagged PSD95 levels are detected. TAP/+, heterozygous for TAP-tagged Arc or PSD95 as indicated; SN, supernatant.