Fig. 4

Experimental validation of differentially expressed exons in Tnnt1 gene. a IGV snapshot of differentially expressed exons identified in Tnnt1 gene. Read coverage of differentially expressed exons 1, 2, 3, 5, and 7 in Tnnt1 gene are shown in all 3 replicates of TSCs and TGCs. Reference gene track is shown at the bottom (gray line) with bars representing the corresponding exons and arrowhead (black) showing the orientation of the gene. b Primer design strategy used for the amplification of differentially expressed exons in Tnnt1 gene. Boxes represent differentially expressed exons (black) in TSCs and exons with no expression (dark gray) in both TSCs and TGCs. Forward primer (top arrow) spanned exons 2 and 3 while reverse primer (bottom arrow) was in exon 7. Connecting line (black) between the boxes represents the intronic region. c Differential amplification of Tnnt1 gene in TSCs and TGCs. Specific exons in Tnnt1 gene and actin were amplified from TSCs and TGCs through PCR. Gel image of the PCR-amplified products of Tnnt1 gene and the positive control (actin) from TSCs and TGCs is shown. PCR amplification from RT-negative (without reverse transcriptase) samples with actin primers represents the negative control. d mRNA expression analysis of differentially expressed exons in Tnnt1 gene by quantitative real-time PCR. Error bars represent SEM of 3 independent biological replicates.