Figure 3.
Mechanisms of cholinergic regulation of hnRNPA2/B1 protein levels. A, No change in ubiquitination status of hnRNPA2/B1 was observed when hnRNPA2/B1 from the hippocampus of VAChT deficient mice (p = 0.3067; control, n = 7; VAChTNkx2.1-Cre-flox/flox mice, n = 6) was immunoprecipitated using anti-hnRNPA2/B1 antibody and probed with anti-ubiquitin antibody. B, Sarkosyl insolubility assay shows no aggregation of hnRNPA2/B1 in VAChTNkx2.1-Cre-flox/flox mice (n = 4). C, Transcript level of hnRNPA2/B1 in the RNA-Seq data set (p = 0.9124, FDR = 1). D, Diagram of the alternative splicing in the 3′UTR of hnRNPA2/B1 transcripts. The FL transcript is predicted to be stable and undergo translation, whereas the NMD-sensitive transcript is not (Bonomi et al., 2013). Primers used to assay this event are show in the schematic. E, Increase in the proportion of NMD+ hnRNPA2/B1 transcripts in the hippocampus of VAChTNkx2.1-Cre-flox/flox mice (n = 4). **p < 0.01.