miRNA sequencing and poly(A) tail length analysis in the hippocampus of Tent2 KO mice. (A) Schematic illustration of Tent2 locus fragment. Position of the 21 bp insert is indicated; the sequence is marked with a yellow box. To facilitate the detection of mutations, a restriction site for BamHI was introduced. STOP codons introduced in the insert are marked with asterisks. (B) Workflow of the experiments presented in panels C–E and on Figure 2, depicting isolation of RNA from the hippocampi and different sequencing approaches. (C–E) Results of miRNA sequencing of Tent2 KO and WT hippocampi (WT n = 3, Tent2 KO n = 5). (C) IsomiR class percentage analysis, showing that 11.1% of isomiRs in WT and 9.8% in Tent2 KO had NT tail at the 3′ end. (D) Significant reduction of adenylation accompanied by an increase in uridylation in the NT tails in Tent2 KO hippocampi. (E) miRNA abundance in Tent2 KO hippocampi when compared to WT.
