Fig 5.

Verification of hnRNP Q-associated mRNAs. (A) Cell lysates of mock-treated (−) or KCl-treated (+) cortical neurons were subjected to immunoprecipitation (IP) using control immunoglobulin (IgG) or anti-hnRNP Q1. The top panel shows immunoblotting of lysates using anti-hnRNP Q1. Coprecipitated RNAs were analyzed by RT-PCR with specific primers; the number of amplification cycles is indicated in parentheses. Input: RT-PCR was performed using mock- and KCl-treated lysates prior to immunoprecipitation. rRNA served as a control. Relative levels of hnRNP Q1-associated RNAs in KCl- versus mock-treated cells are shown; means and standard errors of the means were obtained from three independent experiments. (B) N2A cells were transiently transfected with an empty vector (pEF) or FLAG-hnRNP Q1-expressing vector. The cell lysates were subsequently incubated with ³²P-labeled sense and antisense (con) Cdc42 3′ UTR. After UV cross-linking, the reaction mixtures were subjected to immunoprecipitation using anti-FLAG M2 beads. Lanes 1 to 4 show the input.