Fig. 2: The matrisome controls germ cell number and distal germline structure.

a Bar graphs showing the number of cells within the progenitor zone (PZ—blue) and transition zone (TZ—red) after RNAi. 37 genes affect PZ, 31 affect TZ, and 32 affect both regions. Data are presented as mean values ± SEM. Statistical significance and p values were calculated with respect to control for each RNAi (see Supplementary Figs. 4, 5 and Supplementary Data 5 for details, control values and data distribution for each experiment). b Network showing STRING analysis of association between matrisome genes with validated PZ function and known genes controlling the mitotic cell cycle. Red letter = genes associated with mitotic cell cycle (GO:0000278). Green shade = Genes have direct association in the STRING network. c Representative images of EdU and DAPI (pink) staining showing S phase stages. n per gene knockdown ≥ 6. Scale bar = 5 µm. d Venn diagram showing the most significant matrisome genes for PZ cell number difference (p < 0.0001 based on unpaired t test (for groups of 2) or ordinary one-way ANOVA (for groups of >2)) that control the cell cycle. Pink—total S phase, green—early S phase, yellow—mid S phase and blue—late S phase. e Micrographs showing cytoskeletal structure (phalloidin in yellow) at the distal end. Scale bar = 5 µm. n per gene knockdown ≥5. f Diagram showing the most significant matrisome genes for PZ cell number difference (p < 0.0001 based on unpaired t test (for groups of 2) or ordinary one-way ANOVA (for groups of >2)) that affect cytoskeletal structure. Green—Gene knockdowns with ≥50% germ lines with defective cytoskeleton. g 3D reconstruction of endogenous GLP-1 (blue) and phalloidin (yellow) in the distal germ line. GLP-1 protein distribution and staining phenotype were quantified. Scale bar = 10 µm. n per gene knockdown ≥5. h Diagram showing the most significant matrisome genes for PZ cell number difference (p < 0.0001 based on unpaired t-test (for groups of 2) or ordinary one-way ANOVA (for groups of >2)) and their effect on GLP-1. Green circle- Genes showed an impact on GLP-1 phenotype in ≥50% germ lines. Red circle- Genes showed significant difference in GLP-1 distribution length. n per gene knockdown ≥5. i 3D reconstruction nuclei distal germline (red). Scale bar = 10 µm. i1—The distances between three neighboring nuclei to each nucleus in PZ. j Gene knockdowns with a statistically significant difference in nuclear distribution. n ≥ 5. Supplementary data associated to Fig. 2—Supplementary Figs. 2, 4, 5 and Supplementary Data 5.