Fig. S3

Generation and validation of mstnb LOF line. (a-c) mstnb TALEN designed to target the region after the signal peptide encoded by exon 1 was generated (a), a 10 bp frameshift deletion (mstnb^bns5) was identified (b) and is predicted to cause the formation of truncated Mstnb (c). Red number indicates the last native amino acid before the frameshift mutation; green number indicates the last amino acid before stop codon. (d) RT–qPCR for mstnb expression analysis in mstnb^+/+ and mstnb^-/- adult hearts (n=2 x 3 cardiac ventricles assessed as 2 biological and 2 technical replicates). (e-h) mstnb^+/+ and mstnb^-/- adult hearts, H&E staining of heart sections and higher magnifications of H&E staining showing representative enlarged heart with thickened ventricular wall in mstnb^-/- animals. (i-k) Quantification of ventricle, atrium and eye size in mstnb^+/+ (n=4) and mstnb^-/- (n=5) adults. (l) Quantification of ventricular wall thickness in mstnb^+/+ (n=10) and mstnb^-/- (n=8) adult hearts. (data are mean ± s.e.m., ns: no significant changes observed, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 and ****P ≤ 0.0001 - Student’s t-test, two-tailed). Scale bars, 100 μm.