Fig. 3: Identification of candidate target genes of REPTOR in muscle.
From: REPTOR and CREBRF encode key regulators of muscle energy metabolism
a UMAP plot showing the integration of the datasets of control, dMef2 > REPTOR[ACT] and dMef2>FoxO[ACT] thoraces. b Dot plot representing the scaled expression of three markers of muscle tissue, sallimus, Myofilin and Myosin heavy chain (Mhc). Spalt major is used as marker of indirect flight muscles102. c Schematics of the strategy to identify specific target genes of REPTOR in muscle. All four muscle clusters were included in the analysis. Seventeen genes were identified from the intersection between snRNA-seq muscle clusters and the bulk RNA-seq of dMef2 > REPTOR[ACT]. Three candidate targets of REPTOR were shortlisted based on being upregulated in more than one muscle cluster and having REPTOR/REPTOR-BP DNA binding sites. d–g mRNA levels in thoraces of the candidate targets identified upon muscle-specific induction of REPTOR[ACT] (CG42390 p = 0.017*, Gadd45 p = 0.0035**, RhoGAP68F p = 0.0027**) (d), or knockdown of REPTOR in muscle of Esg>yki[S3A] flies for 12 days (CG42390: Ctrl vs yki[S3A] p < 0.0001****, yki[S3A] vs yki[S3A] REPTOR RNAi p = 0.0084**. RhoGAP68F: Ctrl vs yki[S3A] p < 0.0001****, yki[S3A] vs yki[S3A] REPTOR RNAi p = 0.0002***. Gadd45: Ctrl vs yki[S3A] p < 0.0001****, yki[S3A] vs yki[S3A] REPTOR RNAi p = 0.0002***) (e–g). N = 5–9 biologically independent samples, N = 6–10 thoraces per sample. Data shows boxplots (median and quartiles) with whiskers (minimum to maximum) (d–g). Values were normalized to the mean of control samples (d–g). Statistical analysis was done by using two-tailed t-test with Welch’s correction (d), or one-way ANOVA with Sidak correction test for multiple comparisons (e–g). Source data are provided as a Source Data file.
