Fig S8

Loss of sema4ab reduces il1b and tnfa expression in vivo, but plxnb1a/plxnb1b disruption does not alter the cytokine profile of the lesion site.
(A) A schematic indicating the experimental design for cytokine quantification in situ is shown. (B) il1b:GFP fluorescence intensity reported in transgenic fish is decreased in lesioned sema4ab somatic mutants compared to control gRNA-injected fish at 24 hpl (−1.45 fold-change; Unpaired t test: p = 0.0004). (C) tnfa:EGFP fluorescence intensity reported in transgenic animals is decreased in lesioned sema4ab somatic mutants compared to control injected fish at 24 hpl (−1.49 fold-change; Unpaired t test: p < 0.0001). (D) il1b expression in microglia (apoeb+ cells) detected by HCR shows a reduction of 11% in mean intensity (Unpaired t test: p = 0.0380). (E) A schematic indicating the experimental design for qRT-PCR. (F) qRT-PCR analyses of major pro-inflammatory cytokines (il1b, tnfa, il6) and tgfb3 in plxnb1a/plxnb1b double somatic mutants show no differences to injured control gRNA-injected larvae at 24 hpl (One-sample t test; il1b: p = 0.537; tnfa: p = 0.4840; il6: p = 0.5464; tgfb3: p = 0.687). Each dot for qRT-PCR represents a pool of 50 larvae. β-actin was used as housekeeping gene. Raw data for qRT-PCR can be found in S5 and S6 Data. Error bars show SEM. Scale bars: 50 µm. Data files for graphs available in S3 Data.