Figure 2.

Cardiac Basement Membrane Protein Regulation Differences in wildtype Drosophila. (A) Scatter plot of the MS1 intensity values for each protein. MS1 intensity values (protein abundance) are plotted for w¹¹¹⁸ value on the x-axis and yw value on the y-axis. Colors of points indicate biological function as determined by Software Tool for Rapid Annotation of Proteins (STRAP) analysis and relative percentages in each quadrant are given (ECM - green, Cytoskeleton - blue). Red arrow indicates Pericardin [Prc]. Data represents a mean value of 4 biological replicates of 50 pooled fly hearts for protein analysis. (B) Absolute distance of MS1 intensity values from a zero log₂ fold change or x=y were plotted for each protein in decreasing fashion. 25 proteins with highest calculated distance values are shown in subpanel (grey dashed box) color coded by STRAP analysis (ECM - green, Cytoskeleton - blue) with ECM proteins of interest listed on the x-axis and Laminin A (the cardiac specific laminin subunit within trimer) highlighted with green arrow and Pericardin in red arrow. (C) Representative immunofluorescent images for the indicated genotypes at 1 week for proteins Laminin A (green), Viking (purple), Pericardin (red), and F-actin (white). All fluorescent intensities were normalized to F-actin within region of interest (ROI) selected over bulk of heart tube (top left, white dashed box). Scale bar is 50 µm. (D) Mean pixel intensity for each indicated protein as measured within ROI, normalized to F-actin intensity within ROI, and shown as w¹¹¹⁸ normalized to yw. Mean Factin intensity varied <10% sample to sample between each genotype or with age. n=4 hearts per comparison.