Heat shock proteins and small nucleolar RNAs are dysregulated in a Drosophila model for feline hypertrophic cardiomyopathy

Figures

• 

  Figure 1. Heart rate of D. melanogaster larvae that express feline MYBPC3. Heart rates (BPM) of wild type (WT, w1118/y, w; Chr2/+; UAS-WT-MYBPC3/TinCΔ4-Gal4), A31P (w1118/y, w; Chr2/+; UAS-A31P-MYBPC3/TinCΔ4-Gal4), and R820W (w1118/y, w; Chr2/+; UAS-R820W-MYBPC3/TinCΔ4-Gal4) MYBPC3 alleles expressed in cardiac tissue and the attP2 control (w1118/y, w; Chr2/+; attP2/TinCΔ4-Gal4). We tested all pairwise comparisons among means using the Tukey HSD test. Tukey groupings (A or B) are shown above each bar to indicate significantly different heart-rates. Tukey grouping A has a significantly different heart rate compared to Tukey grouping B.

• 

  Figure 2. Climbing behavior before and after exercise. Climbing behavior before (black bars) and after (grey bars) 1 h of intense exercise of one week old MYBPC3 alleles expressed in cardiac tissue [w1118/y, w; Chr2/+; UAS-WT-MYBPC3/TinCΔ4-Gal4 (WT), w1118/y, w; Chr2/+; UAS-A31P-MYBPC3/TinCΔ4-Gal4 (A31P), w1118/y, w; Chr2/+; UAS-R820W-MYBPC3/TinCΔ4-Gal4 (R820W)] and their control [w1118/y, w; Chr2/+; attP2/TinCΔ4-Gal4 (attP2)]. White bars indicate the difference between pre- and post-exercise. (A) Females. (B) Males. We tested all pairwise comparisons among means using the Tukey HSD test (by sex). Tukey HSD groupings (A–E) are indicated above each bar to indicate means that are significantly different from one another. Results of these statistical analyses are given in Supplementary Table S2.

• 

  Figure 3. Numbers of overlapping or unique differentially expressed genes. The numbers of genes in the Venn diagrams represent significantly differentially expressed genes (FDR < 0.05 and Log2FC > 0.5 or Log2FC < −0.5) from pairwise comparisons of each feline MYBPC3 allele expressed in cardiac tissue and the control genotype (A, top panel) or when the WT nonexercised genotype was compared to the exercised WT, A31P and R820W genotypes (B, bottom panel). Venn diagrams were generated using Venny 2.1 (https://bioinfogp.cnb.csic.es/tools/venny/). The nonexercised conditions are shown in blue and the exercised conditions are in white.

• 

  Figure 4. Heat maps of differentially expressed genes. Heat maps of differentially expressed genes among the MYBPC3 alleles and the attP2 control, expressed in cardiac tissue. The row Z-scores were calculated as Z = (x–μ)/σ where x is gene expression expressed as Log2FC; μ is the mean Log2FC across the samples; and σ is the standard deviation across samples for a given gene. Each row represents a significantly differentially expressed gene. Each column is the MYBPC3 genotype normalized against the attP2 control line. The heat maps were generated using www.heatmapper.ca. (A) Top panel shows heat maps for larvae and the nonexercised and exercised conditions for females and males. (B) Bottom panel shows heat maps comparing the nonexercised WT and exercised WT, A31P and R820W for both females and males.

• 

  Figure 5. Heat maps of differentially expressed small nucleolar RNAs (snoRNAs) between feline MYBPC3 alleles in exercised females. The row Z-scores were calculated as Z = (x–μ)/σ where x is gene expression expressed as Log2FC; μ is the mean Log2FC across the samples; and σ is the standard deviation across samples for a given gene. Each row represents a significantly differentially expressed snoRNA gene and each column is the MYBPC3 genotype normalized against the attP2 control line. The box type, target RNA, organization, and locus were identified using snOPY (http://snoopy.med.miyazaki-u.ac.jp/) (Yoshihama et al. 2013). No other condition exhibited co-regulated differential expression of snoRNAs. The heat maps were generated using www.heatmapper.ca.

• 

  Figure 6. Heat maps of differentially expressed Heat shock proteins (Hsps) between feline MYBPC3 alleles. The row Z-scores were calculated as Z = (x–μ)/σ where x is gene expression expressed as Log2FC; μ is the mean Log2FC across the samples; and σ is the standard deviation across samples for a given gene. Each row represents a significantly differentially expressed Hsp or Hsc gene. Each column is the MYBPC3 genotype normalized against the attP2 control line. The heat maps were generated using www.heatmapper.ca.