Extended Data Fig. 2: Natural Aging Downregulates LamC and LamB but does not affect their localization.
(A) MA plot of all genes from 1- and 5-week adult w1118 fly hearts showing log2 fold change (FC) and mean normalized expression counts. Data is shown in black for genes −1.25 < FC < 1.25 (dashed lines) or p-adj > 0.05. Open circles represent genes that do not map to a nuclear ontological term. Green and purple data represent DEGs that are up- or downregulated in 5-week adult flies, respectively. (B) Cellular component and molecular function ontological terms for genes associated with the nuclear envelope GO term, organized by their elimination pruning p-value. (C) Representative images of 1 and 5 week w1118 fly heart nuclei stained for DNA, and LamC (green) and LamB (purple) mRNA transcripts. Scale bar, 5 μm. (D) Left plot shows area percentage occupied by mRNA transcripts per cardiomyocyte in 1- and 5-week-old adult w1118 fly hearts. Right plot normalizes data to mean area at 1 week for each transcript. n = 49, 36, 49, and 36 nuclei from w1118 flies at 1 and 5 weeks for LamC and LamB, respectively. (E) Corrected total nuclear fluorescence (CTNF) of 1, 3, and 5 week yw flies for LamC (top) and LamB (bottom). n = 94, 106, and 133 nuclei for LamC and n = 173, 115, and 90 nuclei for LamB for 1, 3, and 5 week adults, respectively. (F) Image showing a representative nucleus with multiple lines radiating out from its centroid (left) to create line plots that are averaged into a single radial profile of the fluorescent intensity (right). Lower panel, the ratio of edge to center intensity is plotted. n = 72, 63, 73, and 33 nuclei from w1118 flies at 1 and 5 weeks for LamC and LamB, respectively. n = 51, 87, 77, and 42 nuclei from yw flies at 1 and 5 weeks for LamC and LamB, respectively. *p < 0.05, **p < 10−2, ***p < 10−3, and ****p < 10-4 by one-way ANOVA with Tukey multiple comparisons test. Bars in (D) refer to min to max, with median and 25th and 75th interquartile range and error bars in (E-F) refer to mean + /- SD.
