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Fig. 6 | Molecular Brain

Fig. 6

From: Neuronal activity regulates alternative exon usage

Fig. 6

Activity dependent alternative splicing of Inhba. a Inhba is encoded by 5 exons (numbered, yellow boxes, not to scale). Transcript 1 and 2 (green) were described before. Transcript 3 and 4 (white) were identified in this study. Two additional transcribed exons encoding an lncRNA (brown), one upstream and one between exon 4 and 5 of Inhba are shown. b Visualization of the microarray data obtained for Inhba. The plot shows the log2 ratio calculated from the mean normalized intensities of transformed expression values over the course of probesets per exon. Each green line represents expression kinetics of one time point after seizure onset. The blue boxes represent the probesets per exon present on the microarrays. c Validation of Inhba variant expression in hippocampus of control mice or 1, 4 or 8 h after seizure onset by RNA sequencing. Shown are coverage plots (blue) from paired-end reads for the four sample groups. The reference gene tracks are depicted below (black, Inhba and brown, lncRNA). The red circle highlights expression of one exon of the lncRNA. d Sashimi plot of RNA sequencing data for cda. Numbers indicate the counts of RNA sequencing reads that span the respective exon junctions. e, f Validation of splicing events by RT-PCR using indicated exon-spanning primer pairs (arrows). The respective exons are indicated by numbers and in green for Inhba (e) and in brown for the lncRNA (f). Primers were used to assess expression of exons in total RNA of hippocampi of control mice (−) and of mice sacrificed 4 h after onset of seizures (+). Note that in the left panel two products were amplified, one corresponding in size to amplification of exon 2–4 and one to amplification of exon 2 and 4 and excluding exon 3

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