wnt β catenin agonist skl2001  (MedChemExpress)

Journal: Communications Biology

Article Title: Wnt/FGF20 signaling axis promotes migration of proximal blastema cell during tail regeneration of Gekko japonicus

doi: 10.1038/s42003-025-09360-6

Figure Lengend Snippet: a EdU signals (red)of proliferating cells after 12 h and 24 h of SKL2001 (40 µM) treatment. Scale bar represents 75 µm. b Quantification of EdU-positive cells. The data showed that proliferation of cultured blastema cells was decreased by 37.8% at 12 h and 55% at 24 h following treatment with SKL2001. Data were analyzed using two-way ANOVA followed by Bonferroni’s post hoc test and are presented as mean ± SEM, n = 6. The P values are shown on the panel. NC negative control.

Article Snippet: The cultured cells were treated with the Wnt/β-catenin signaling agonist SKL2001 (40 μM, Cat# S8320, Selleck Chemicals) for 24 hours prior to assays.

Techniques: Cell Culture, Negative Control

Journal: Communications Biology

Article Title: Wnt/FGF20 signaling axis promotes migration of proximal blastema cell during tail regeneration of Gekko japonicus

doi: 10.1038/s42003-025-09360-6

Figure Lengend Snippet: a qRT-PCR results showed that SKL2001 treatment reduced fgfr4 mRNA levels by 31.4% in cultured blastema cells. Data were analyzed using an unpaired Student’s t-test and are presented as mean ± SEM, n = 5. The P value is shown on the panel . b qRT-PCR analysis of fgfr4 mRNA levels in the 1 mm tip portion of the distal blastema on days 8, 10, 12, 14 post-autotomy. Data were analyzed using one-way ANOVA followed by Dunnett’s post hoc test and are presented as mean ± SEM, n = 5. The P value is shown on the panel. c Representative immunofluorescence and EdU staining images showed FGFR4 (green) expression and proliferating cells (EdU, violet) in regenerated tissues on days 8, 10, 12, and 14 post-autotomy. The region between the solid and dotted lines represent WE. Scale bars represent 50 µm. d Normalized FGFR4 fluorescence intensity of z1-z5 on day 14. Integrated density was divided by the corresponding cell area and expressed as fluorescence intensity per unit area (arbitrary units, a. u.). The mean value of the control group was set to 1. Data were analyzed using one-way ANOVA followed by Dunnett’s post hoc test and are presented as mean ± SEM, n = 5. The P value is shown on the panel. e Schematic timeline for DMSO/IWP4 injection and sample collection. f qRT-PCR results showed a 67.2% increase in the mRNA levels of fgfr4 in the blastema after Wnt/β-catenin inhibition. Data were analyzed using an unpaired Student’s t -test and are presented as mean ± SEM, n = 5. The P value is shown on the panel. WE, wound epithelium. NC, negative control. qRT-PCR, quantitative reverse transcription.

Article Snippet: The cultured cells were treated with the Wnt/β-catenin signaling agonist SKL2001 (40 μM, Cat# S8320, Selleck Chemicals) for 24 hours prior to assays.

Techniques: Quantitative RT-PCR, Cell Culture, Immunofluorescence, Staining, Expressing, Fluorescence, Control, Injection, Inhibition, Negative Control, Reverse Transcription

Journal: Communications Biology

Article Title: Wnt/FGF20 signaling axis promotes migration of proximal blastema cell during tail regeneration of Gekko japonicus

doi: 10.1038/s42003-025-09360-6

Figure Lengend Snippet: a Schematic timeline for DMSO/BLU9931 injection and sample collection. b The regeneration stage were observed on days 8 and 12 post-autotomy. The results of statistical analysis showed that FGFR4 inhibitor BLU9931 delayed tail regeneration. Data were analyzed by χ 2 test. The P values are shown on the panel. c EdU staining (red) of proliferating cells in DMSO group and BLU9931-treatment group. d Quantification of EdU-positive cells in WE and blastema. In DMSO group, the percentage of EdU-positive cells in WE = (56.9 ± 3.5) %, blastema = (48.3 ± 4.2) %. In BLU9931 group, the percentage of EdU-positive cells in WE = (24.7 ± 2.5) %, blastema = (25.9 ± 3.2) %. The results indicated that BLU9931 treatment suppressed cell proliferation in both wound epithelium and the blastema. Data were analyzed using two-way ANOVA followed by Bonferroni’s post hoc test and are presented as mean ± SEM, n = 5. The P values are shown on the panel. e , f EdU assay of proliferating cells after treatment with Wnt agonist SKL2001 combined with fgfr4 over-expression (OE- fgfr4 ) in vitro and quantification of EdU-positive cells showed that fgfr4 over-expression alleviated the inhibitory effect of Wnt signaling on cell proliferation. Data were analyzed using two-way ANOVA followed by Bonferroni’s post hoc test and are presented as mean ± SEM, n = 5, The P values are shown on the panel. WE, wound epithelium.

Article Snippet: The cultured cells were treated with the Wnt/β-catenin signaling agonist SKL2001 (40 μM, Cat# S8320, Selleck Chemicals) for 24 hours prior to assays.

Techniques: Injection, Staining, EdU Assay, Over Expression, In Vitro

Journal: Communications Biology

Article Title: Wnt/FGF20 signaling axis promotes migration of proximal blastema cell during tail regeneration of Gekko japonicus

doi: 10.1038/s42003-025-09360-6

Figure Lengend Snippet: a Schematic of the co-culture system. b EdU signal (red) of proliferating cells in the co-culture proliferation system after 12 h and 24 h. Blastema cells treated with SKL2001 were cultured in the upper chamber, while untreated cells were cultured in the lower chamber. Red represents EdU, blue color indicates nucleus. Scale bar represents 75 µm. c Quantification of EdU-positive cells indicated that Wnt-activated cells did not affect the proliferation of unactivated cells. Data were analyzed using two-way ANOVA followed by Bonferroni’s post hoc test and are presented as mean ± SEM, n = 6, The P values are shown on the panel. d Co-culture transwell migration assay, where blastema cells treated with SKL2001 were cultured in the lower chamber and untreated cells in the upper chamber. Blue cells represent the cells that migrated to the lower surface of the upper chamber. Scale bar represents 75 µm. e Quantification of migrating cells showed that Wnt-activated cells led to a 79.7% increase in the migration of un-activated cells. Data were analyzed using an unpaired Student’s t -test and are presented as mean ± SEM, n = 6. The P value is shown on the panel. NC negative control.

Article Snippet: The cultured cells were treated with the Wnt/β-catenin signaling agonist SKL2001 (40 μM, Cat# S8320, Selleck Chemicals) for 24 hours prior to assays.

Techniques: Co-Culture Assay, Cell Culture, Transwell Migration Assay, Migration, Negative Control

Journal: Communications Biology

Article Title: Wnt/FGF20 signaling axis promotes migration of proximal blastema cell during tail regeneration of Gekko japonicus

doi: 10.1038/s42003-025-09360-6

Figure Lengend Snippet: a qRT-PCR results showed that fgf20 mRNA levels were increased by 4.32-fold after SKL2001 treatment in cultured blastema cells. Data were analyzed using an unpaired Student’s t -test and are presented as mean ± SEM, n = 5. The P value is shown on the panel. b Prediction of potential binding sites for TCF/LEF-1 in the fgf20 promoter region using JASPAR and AnimalTFDB v4.0. c Luciferase reporter assay showed that β-catenin significantly enhanced fgf20 promoter activity. Data were analyzed using an unpaired Student’s t -test and are presented as mean ± SEM, n = 5, The P value is shown on the panel. d Immunofluorescence analysis of FGF20 expression in regenerated tissues on days 8, 10, 12 and 14 post-autotomy. The region between the solid and dotted lines represents the WE. Enlarged views of dashed boxed areas are shown in the bottom panel. Scale bars represent 200 µm (top panel) and 50 µm (bottom panel). The region between the solid and dotted lines represent the WE. e qRT-PCR analysis of relative mRNA levels of fgf20 on day 8, 10, 12 and 14 post-autotomy. The fgf20 mRNA level on day 8 was normalized as 1, and (day 10) = 1.96 ± 0.43, (day 12) = 3.59 ± 0.29, (day 14) = 9.17 ± 0.71. f qRT-PCR results showed an 83.8% decrease in the mRNA levels of fgf20 in the blastema on day 12 after Wnt/β-catenin inhibition. Data were analyzed using an unpaired Student’s t -test and are presented as mean ± SEM, n = 5. The P value is shown on the panel. NC negative control. qRT-PCR quantitative reverse transcription. WE wound epithelium.

Article Snippet: The cultured cells were treated with the Wnt/β-catenin signaling agonist SKL2001 (40 μM, Cat# S8320, Selleck Chemicals) for 24 hours prior to assays.

Techniques: Quantitative RT-PCR, Cell Culture, Binding Assay, Luciferase, Reporter Assay, Activity Assay, Immunofluorescence, Expressing, Inhibition, Negative Control, Reverse Transcription

Journal: Communications Biology

Article Title: Wnt/FGF20 signaling axis promotes migration of proximal blastema cell during tail regeneration of Gekko japonicus

doi: 10.1038/s42003-025-09360-6

Figure Lengend Snippet: a Schematic of the co-culture system. b EdU signal of proliferating cells in the co-culture proliferation system. fgf20 -overexpressing blastema cells were cultured in the upper chamber, while untreated cells were cultured in the lower chamber. c Quantification of EdU-positive cells showed that fgf20 -overexpressing cells do not affect the proliferation of unactivated cells. Data were analyzed using an unpaired Student’s t -test and are presented as mean ± SEM, n = 6, The P value is shown on the panel. d Co-culture transwell migration assay, where fgf20 -overexpressing blastema cells were cultured in the lower chamber, and untreated cells were cultured in the upper chamber. e Quantification of migrating cells showed that fgf20 -overexpressing cells led to a 2.47-fold increase in the migration of unactivated cells. Data were analyzed using an unpaired Student’s t -test and are presented as mean ± SEM, n = 6, The P value is shown on the panel. f Co-culture transwell migration assay. The blastema cells were cultured in upper chamber and fgf20 knockdown blastema cells, in the presence or absence of SKL2001, cultured in the lower chamber. Scale bar represents 75 µm ( b , d , f ). g Quantification of migration showed that fgf20 knockdown significantly reduced migration of untreated blastema cells and attenuated the promotive effect of Wnt signaling on cell migration. Data were analyzed using two-way ANOVA followed by Bonferroni’s post hoc test and are presented as mean ± SEM, n = 5, The P values are shown on the panel.

Article Snippet: The cultured cells were treated with the Wnt/β-catenin signaling agonist SKL2001 (40 μM, Cat# S8320, Selleck Chemicals) for 24 hours prior to assays.

Techniques: Co-Culture Assay, Cell Culture, Transwell Migration Assay, Migration, Knockdown