a Schematic of key peroxisome proteins and functions. b–e Peroxisomes (SKL-GFP) and analysis and quantification of peroxisome density in PCs and SCs. f–h Peroxisomes (SKL-GFP) interacting with lipid droplets (LipidTox) in PCs via putative pexapodia, along with quantification. i Peroxisomes (SKL-GFP), lipid droplets (LipidTox) and mitochondria (MitoView) in PCs, high magnification examples of tight inter-organelle associations. j Expression of key peroxisome proteins in PCs and SCs from publicly available snRNA-Seq data (Fly Cell Atlas). Secretory activity (k), bioenergetic output (l) and LD density (m–o) in Pex3 or Pex16 RNAi tubules (driven by CapaR-gal4). Data represented as box and whisker plots (lower and upper hinges correspond to the first and third quartiles, median line within the box, whiskers extend from the hinge to the largest/smallest value, at most 1.5* interquartile range of the hinge) with all data from MpT cells (SCs or PCs, e), MpT main segment sections (l, m) or secretion of individual kidneys (k) shown as overlaid points. NS Not Significant, **P < 0.01, ***P < 0.001 (unpaired two-tailed t-tests, ANOVA followed by Tukey multiple comparisons test where appropriate or Wilcoxon test with FDR correction). p values: (e) p < 0.0001, (k) control vs Pex16 p = 0.00288, control vs Pex3 p = 0.000303, (l) p = 0.000484, (m) p < 0.0001. p values for (j) displayed on the Figure. For analysis of fluorescent reporters/dyes, two images of different sections of the MpT main segment per fly were imaged. All images representative of >5 tubules. All images are maximum z projections. e
n = 12 PCs (from 6 tubules) and 13 SCs (from 5 tubules), (h) n = 7 tubules, (k) n = 28 control, 25 Pex16 and 19 Pex3 tubules, (l) n = 12 tubules per condition, (m) n = 11 control and 12 RNAi tubules. Source data are provided as a Source Data file.