Fig. 4: mmpReg promotes blastema formation.
a qPCR for the RNAi-mediated downregulation of mmpReg transcripts at 24 hpa (n = 4, biologically independent sample pools, p = 0.00732). Error bars indicate S.E.M. b, c Representative images of animals at 6 hpa. Bright-field images (b). The nuclei were stained with DAPI (c). d, e RNAi of mmpReg reduced the number of cells in both the anterior blastema (GFP RNAi (n = 22), mmpReg RNAi (n = 30), p = 0.00109) (d) and posterior blastema (GFP RNAi (n = 22), mmpReg RNAi (n = 29), p = 2.06 × 10−4) (e). Representative of two independent experiments (d, e). f, g Representative images of animals at 24 hpa. Bright-field images (f). The nuclei were stained with DAPI (g). h, i RNAi of mmpReg reduced the number of cells in the anterior blastema (GFP RNAi (n = 20), mmpReg RNAi (n = 32), p = 9.1 × 10−5) (h) and posterior blastema (GFP RNAi (n = 20), mmpReg RNAi (n = 31), p = 0.00165) (i). Representative of two independent experiments (h, i). j–m mmpReg RNAi resulted in a decrease in the blastema volume at 6 hpa (GFP RNAi (n = 22), mmpReg RNAi (n = 20), p = 4.8 × 10−6) (k) or 24 hpa (GFP RNAi (n = 24), mmpReg RNAi (n = 29), p = 2.7 × 10−4) (m). Representative images are shown (RNAi 6 hpa (j) or 24 hpa (l)). n, o MMP inhibitor (MMP2/MMP9 inhibitor I, N-([1,1’-biphenyl]−4-ylsulfonyl)-D-phenylalanine, CAS#:193807−58-8) resulted in decreased blastema volume at 24 hpa (DMSO (n = 22), inhibitor (n = 24), p = 1.4 × 10−5) (o). Representative images are shown (MMP inhibitor (n)). Data was combined from two independent experiments (k, m, o). t-test, *p < 0.05, **p < 0.01, ***p < 0.001(a, d, e, h, i, k, m, o). Error bars: mean ± SD (d, e, h, i, k, m, o). Scale bars represent 250 µm (c, g, j, l, n). Dashed lines indicate amputation sites (b, c, f, g, j, l, n). Dotted lines indicate the outline of regenerating worms (c, g). Ventral views (b, c, f, g). Sagittal sections (j, l, n). Source data are provided as a Source Data file.
