(A) Western blots show that Rab4a protein (but not Sec23a) is increased in KO neurons relative to controls in three independent neuron culture isolation experiments (i1, i2, i3), as detected by a mouse polyclonal antibody (SIGMA). (B) Densitometry shows significant increase in Rab4a levels in KO neurons vs. controls. (C–F) Neurons were transduced with lentivirus to overexpress NT-shRNA or RBM5 targeting shRNA and protein extracts were harvested between DIV3-DIV12. Two biological replicates (n=1 and n=2) were prepared/harvested from a single neuron culture isolation experiment. (C and D) Age course samples were probed with anti-RBM5, anti-Rab4 (SIGMA, Mouse Polyclonal), and anti-α-Tubulin antibodies. (E and F) Age course samples were probed with anti-RBM5, anti-Rab4 (Abcam, Rabbit Monoclonal), and anti-α-Tubulin antibodies. Data were analyzed by unpaired T-test. Data were significant at p <.05 (*). Graphs show mean + SEM.