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. 2019 Jul 25;4(14):e128643. doi: 10.1172/jci.insight.128643

Figure 4. EHTs homozygous for R451G maintain normal levels of DSP mRNA.

Figure 4

(A) Genotyping of the control cell line (WT/WT) and the same cell line after CRISPR/Cas9 manipulation to introduce the DSP R451G mutation (ENST00000379802.8; c.1596CGT>GGT; Chr. 6:7568521, GRCh38.p12) at both alleles (R451G/R451G). (B) RT-qPCR was performed on engineered heart tissues formed from WT/WT and R451G/R451G cardiomyocytes in order to probe for DSP mRNA abundance. Several different primer pairs covering different exon junctions indicate that similar levels of DSP mRNA were expressed in both cell lines and that the introduction of c.1596CGT>GGT did not affect mRNA stability or result in alternative splicing (2-tailed unpaired t test with Bonferroni correction; n = 3). Blue and orange arrows represent regions targeted by forward and reverse primer pairs. Cycle thresholds were normalized to TNNT2 (cardiac troponin T).