Journal: Science Advances
Article Title: Divergent stem cell mechanisms govern the primary body axis and appendage regeneration in the axolotl
doi: 10.1126/sciadv.adx5697
Figure Lengend Snippet: ( A ) UMAP visualization of refined subclusters within the regeneration active zone. MuSC, IM1, and IM2 are partitioned into nine subclusters based on shared gene expression programs between progenitors and their progeny. Each cell type, e.g., IM2, is subdivided into subclusters committed to myoblasts (IM2_Myo), fibroblast 2 (IM2_Fib2), or uncommitted (IM2_UC). ( B ) Heatmap showing the scores of predicted pathway activity in nine defined subclusters from (A), combined with Fib1, Fib2, and Myo. ( C and D ) SB-431542 inhibits the production of fibroblasts during tail regeneration. Images of CHERRY fluorescence of 26-day tail regenerates treated with SB-431542 (D) and control (C) ( n = 3 each). ( E and F ) Quantification of CHERRY signal area in the fin and regenerated fin length following SB-431542 treatment ( n = 3). dpt, days posttreatment. Error bars, SEM; statistical analysis was conducted using Student’s t test (E) and one-way analysis of variance (ANOVA) (F); ** P < 0.01, **** P < 0.0001; ns, not significant. ( G ) p-SMAD2 and SMAD2 immunoblots of regenerated tails treated with SB-431542 and control ( n = 3 each). Glyceraldehyde-3-phosphate dehydrogenase immunoblot, loading control. ( H and I ) Quantification of signal intensity from p-SMAD2 and SMAD2 immunoblots in (G) ( n = 3 each). Student’s t test; ** P < 0.01. ( J ) Boxplot of the scores of endogenous TGF-β activity in MuSCs. Note that scores are attenuated in IM2 committed to myoblasts (IM2_Myo), in contrast, elevated in IMs to fibroblasts (IM1_Fib1, IM2_Fib2), compared to that in MuSCs at 0 days postamputation (dpa). ( K and L ) Cell fate trajectory predictions following in silico perturbation of TGF-β signaling. Activation of TGF-β diverts cells from muscle-related lineages to fibroblast-related lineages (K), whereas its suppression diverts cells from fibroblast-related lineages to muscle-related lineages (L). Rectangles in (C) and (D) are shown at higher magnification. Scale bars, 1 mm [(C) and (D)]. See also fig. S20.
Article Snippet: SB-431542 (MedChemExpress, HY-10431) and IWR-1 (MedChemExpress, HY-12238) were dissolved in DMSO at 30 mM stock solutions, and U0126 (MedChemExpress, HY-12031) was dissolved in absolute ethanol.
Techniques: Gene Expression, Activity Assay, Fluorescence, Control, Western Blot, In Silico, Activation Assay
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MedChemExpress manufactures this product 
![( A ) UMAP visualization of refined subclusters within the regeneration active zone. MuSC, IM1, and IM2 are partitioned into nine subclusters based on shared gene expression programs between progenitors and their progeny. Each cell type, e.g., IM2, is subdivided into subclusters committed to myoblasts (IM2_Myo), fibroblast 2 (IM2_Fib2), or uncommitted (IM2_UC). ( B ) Heatmap showing the scores of predicted pathway activity in nine defined subclusters from (A), combined with Fib1, Fib2, and Myo. ( C and D <t>)</t> <t>SB-431542</t> inhibits the production of fibroblasts during tail regeneration. Images of CHERRY fluorescence of 26-day tail regenerates treated with SB-431542 (D) and control (C) ( n = 3 each). ( E and F ) Quantification of CHERRY signal area in the fin and regenerated fin length following SB-431542 treatment ( n = 3). dpt, days posttreatment. Error bars, SEM; statistical analysis was conducted using Student’s t test (E) and one-way analysis of variance (ANOVA) (F); ** P < 0.01, **** P < 0.0001; ns, not significant. ( G ) p-SMAD2 and SMAD2 immunoblots of regenerated tails treated with SB-431542 and control ( n = 3 each). Glyceraldehyde-3-phosphate dehydrogenase immunoblot, loading control. ( H and I ) Quantification of signal intensity from p-SMAD2 and SMAD2 immunoblots in (G) ( n = 3 each). Student’s t test; ** P < 0.01. ( J ) Boxplot of the scores of endogenous TGF-β activity in MuSCs. Note that scores are attenuated in IM2 committed to myoblasts (IM2_Myo), in contrast, elevated in IMs to fibroblasts (IM1_Fib1, IM2_Fib2), compared to that in MuSCs at 0 days postamputation (dpa). ( K and L ) Cell fate trajectory predictions following in silico perturbation of TGF-β signaling. Activation of TGF-β diverts cells from muscle-related lineages to fibroblast-related lineages (K), whereas its suppression diverts cells from fibroblast-related lineages to muscle-related lineages (L). Rectangles in (C) and (D) are shown at higher magnification. Scale bars, 1 mm [(C) and (D)]. See also fig. S20.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_0522/pmc12880522/pmc12880522__sciadv.adx5697-f7.jpg)