Fig. 4
From: Intronic tRNAs of mitochondrial origin regulate constitutive and alternative splicing
NimtRNAs increase downstream exon inclusion in a structure- and position-dependent manner. a Single nimtRNAs were analyzed for their effects on reporter host gene mRNA abundance compared to a scrambled control (scrbl). b Different domains of nimtRNATyr were deleted, i.e., the D-arm (delD), the T-arm (delT), the acceptor stem (delAcc), or the anticodon arm (delAnti), respectively. Alternatively, nimtRNATyr was exchanged for a scrambled sequence (scrbl), or inserted into the splicing reporter in reverse-complementary orientation (r.-c.). c NimtRNATyr was integrated into different locations within the intron of the Low0-eGFP splicing reporter. Respective host gene mRNA abundances were assessed by RT-qPCR, error bars represent the standard deviation from three independent experiments. Normalization was performed relative to a co-transfected reporter control, i.e., Dsred2express. d The nimtRNATyr was inserted into different locations within the introns of the alternative splicing reporter and used for transient transfection experiments. e Alternative splicing isoforms were analyzed by RT-PCR and subsequent gel electrophoresis. PSI (percent spliced in) including standard deviation was quantified from three independent sets of experiments. f, g RT-PCR analysis was performed to detect alternative splicing isoforms of constructs containing different mtRNAs or nimtRNAs within the first intron of the Designer Exon. PSI including standard deviation was quantified from three independent sets of experiments