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Fig. 6
(A) Experimental design used to inhibit glycolysis. Fish are administered, via IP injection, with control (PBS) or 2DG every 12 hr, from fin amputation (0 hpa) until 48 hpa. (B) Schematic representation of the distribution of OBs subtypes along the blastema. (C–D) Representative cryosection images of 48 hpa osx:mCherry (green) caudal fins immunostained for Runx2 (magenta) and counterstained with DAPI (blue), in fish treated with (C,C’) PBS and (D,D’) 2DG. Dashed boxes represent magnified panels in C’ and D’. Arrows indicate Runx2 +Osx- pre-OBs. Dashed arrows indicate Runx2 +Osx + immature OBs. (E–F) Total number of (E) Run2 +Osx and (F) Runx2 +Osx + subtypes in 48 hpa fins treated with PBS or 2DG (PBS = 27, 2DG = 25 cryosections). (G) Percentage of Runx2+/Osx- and Runx2 +Osx + subtypes in 48 hpa fins treated with PBS or 2DG. (H-I’) Representative cryosection images of 48 hpa osx:mCherry (orange) caudal fins immunostained for Runx2 (magenta), EdU (green) and counterstained with DAPI (blue), in fish treated with (H,H’) PBS and (I,I’) 2DG. Dashed boxes represent magnified panels in H’ and I’. Arrows indicate proliferative Edu +Runx2+Osx- pre-OBs. Dashed arrows indicate proliferative Edu +Runx2+Osx + immature OBs. Arrowheads indicate amputation plane. Scale bar represents 100 µm and 20 µm in magnified panels. (J) Total number of Runx2 +Osx + and Runx2 +Osx- proliferative OBs subtypes at 48 hpa fins, treated with PBS or 2DG (PBS = 28, 2DG = 27 cryosections). (K) Percentage of proliferative Runx2 +Osx + and Runx2 +Osx OBs subtypes in 48 hpa caudal fins , treated with PBS or 2DG. E: epidermis; B: bone; M: mesenchyme. For all graphs, statistical analysis corresponds to Mann-Whitney and Mean ± SD are displayed. ns: not significant; **p<0.0001. See Figure 6—source data 1.