Extended Data Fig. 6: Effects of LamC and Myogenic Transcription Factor Overexpression.
For panels A-D and as outlined in Fig. 6a, flies were reared at 18 °C and shifted to 25 °C upon eclosure. Nuclear area (A) and CTNF (B) for hearts overexpressing LacZ (control) and LamC from 1 to 7 weeks. n = 31, 41, 23, 30, 26, and 31 cells (A, left to right) and n = 31, 31, 24, 29, 30, and 40 cells (B, left to right). (C) Diastolic and (D) systolic diameters of LacZ and Hand OE flies at 1, 4, and 7 weeks of adulthood. (E) Representative images (left) for nuclei from flies at 18 °C and 29 °C expressing GFPNLS transgene or LamC OE from HandGal,tubGal80ts;tubGal80ts based upon regime described in S5H. Scale bar is 5 μm. Plots of (F) projected nuclear area, (G) circularity, and (H) and corrected total nuclear fluorescence (CTNF) of LamC protein as a function of temperature and transgene expression. n = 39, 40, 46, and 41 nuclei (left to right). (I) For GFPNLS and LamC OE at 18 °C and 29 °C, representative images of LamC mRNA transcripts. Scale bar is 5 μm. (J) Quantification of LamC transcript area per CM in GFPNLS and LamC OE hearts, at 18 °C and 29 °C. n = 24, 36, 20 and 40 CMs, left to right. Diastolic (K) and systolic (L) diameters, and (M) fractional shortening for GFPNLS and LamC OE at 18 °C and 29 °C. n = 32, 19, 30 and 26 heart tubes, left to right. (N). A plot for each transcription factor is shown and quantifies the per cell, percentage area for each transcript. For all transcripts, n = 50, 56, 49 and 35 cells, left to right. (O) Representative images for tin (pink), Hand (brown), and H15 (blue) transcripts in GFPNLS and LamC OE at 18 °C and 29 °C. Scale bar is 5 μm. *p < 0.05, **p < 10−2, ***p < 10−3, and ****p < 10-4 by two-way ANOVA with Sidaks multiple comparisons test (A-D) and one-way ANOVA with Kruskal-Wallis test and Dunn’s comparisons test (F-N). Error bars in (A, B, F-H) refer to mean + /- SD and all box plots display min to max, with median and 25th and 75th interquartile range.
