Journal: Oncotarget

Article Title: The posterior HOXD locus: Its contribution to phenotype and malignancy of Ewing sarcoma.

doi: 10.18632/oncotarget.9702

Figure Lengend Snippet: Figure 2: DKK2 increases HOXD gene expression through WNT signaling pathway. (A) Increased HOXD10, HOXD11 or HOXD13 expression (bottom) after transfection with cDNA encoding human DKK2 (top) in ES lines A673, MHH-ES1, SK-ES1 or NB line SH-SY5Y, respectively. Results of qRT-PCRs are shown. Data are mean ± SEM; t-test. (B) Analysis of HOXD gene expression in A673, SK-N-MC and TC-71 cells after incubation with recombinant human WNT3a, WNT5a or WNT11 and the combination of all three ligands. RNA was isolated after 1, 3, 6 and 12 h and the time point with the highest increase was shown. Data are mean ± SEM; t-test. (C) Expression of endogenous WNT3a, WNT5a and WNT11 mRNA in three ES cell lines (A673, SK-N-MC and TC-71) analyzed by qRT-PCR. Data are mean ± SEM; t-test.

Article Snippet: To analyze a possible involvement of WNT signaling triggered by WNT ligands on HOXD expression, we incubated A673, SK-N-MC and TC-71 with two different concentrations of recombinant human WNT3a (5036-WN, 0.1μg/ml or 0.3μg/ml), WNT5a (645-WN, 0.1μg/ml or 0.3μg/ml) or WNT11 (6179-WN, 0.6μg/ml or 1.2mg/ml; all R&D Systems).

Techniques: Gene Expression, Expressing, Transfection, Incubation, Recombinant, Isolation, Quantitative RT-PCR

Journal: Frontiers in Cell and Developmental Biology

Article Title: Cecal growth factors promote enteric neurosphere formation and hindgut colonization in the avian model

doi: 10.3389/fcell.2025.1681844

Figure Lengend Snippet: Isolation, characterization and differentiation of chicken enteric neurospheres. Schematic figure of avian enteric neurosphere generation (A) . Brightfield images show examples of neurospheres generated from E8 ENSCs that emerged 1, 3, and 7 days in GDNF-containing medium (B–D) . Representative immunofluorescence images of 7-day-old neurospheres generated from E8 chicken intestine (E–G) . Samples were cultured under three different conditions: Exp.1: control medium ( (E) , CTRL), Exp. 2: GDNF containing cell culture media ( (F) , GDNF), and Exp.3: GWEN ( (G) , ceca-derived mesenchymal growth factors: GDNF, WNT11, ET-3, Noggin) supplemented media. Immunofluorescence of cross-sections confirms the presence of neurons expressing TUJ1, and neural crest cells expressing PHOX2B. Compared to CTRL and GDNF-only conditions, GWEN treatment resulted in a robust increase in TUJ1+/PHOX2B+ cell aggregates on the surface of the neurosphere and single Phox2b+ cells in the center of the aggregates. The inset shows a magnified view of a PHOX2B+/TUJ1+ aggregate, indicating enhanced differentiation of neural crest derivatives (E–G) . Scale bar = 100 µm. CTRL, control; NS, neurosphere.

Article Snippet: To assess the effect of ceca-specific mesenchymal factors on enteric neural stem cells, the culture medium was further supplemented with 10 ng/mL glial cell line-derived neurotrophic factor (GDNF; R&D Systems, 212-GD-010), 200 ng/mL WNT11 (Origene, TP761904), 250 ng/mL endothelin-3 (ET-3; R&D Systems, 1,162/100U), and 100 ng/mL Noggin (R&D Systems, 6997-NG-025).

Techniques: Isolation, Generated, Immunofluorescence, Cell Culture, Control, Derivative Assay, Expressing

Journal: Cells

Article Title: Comparison of Differentiation Pattern and WNT/SHH Signaling in Pluripotent Stem Cells Cultured under Different Conditions

doi: 10.3390/cells10102743

Figure Lengend Snippet: WNT11 treatment and analysis of WNT11 receptors in undifferentiated and differentiating ESCs. ( A ) Expression of Nanog , Pax6 , Gata4 and T in WNT11-treated and control ESCs, EBs and EBOs cultured in medium supplemented with SR. Cells were collected after 48h of WNT11 treatment. ( B ) Expression of WNT11 receptors: Fzd5 and Fzd7 . ( C ) FACS analysis: proportion of undifferentiated ESCs and cells obtained after disaggregation of EB2, EB5, and EB7 synthesizing FZD5 or FZD7. Data presented as means of three independent experiments with standard deviations. Representative histograms present proportion of EB5-derived cells synthesizing FZD5 or FZD7 or cells incubated with IgG only (negative control). Cells were cultured in medium supplemented either with SR or FCS. For qPCR analyzes ( A , B ) β-actin was used as a reference gene. RQ = 1 for the level of gene expression detected in a 13.5-day-old mouse embryo. The average ΔCq for reference sample is shown on each graph. Data are presented as means of three independent experiments with standard deviations. * p < 0.05; Red color indicates differences between undifferentiated ESCs and EBs or EBOs; black color indicates differences between cells cultured in medium supplemented either with FCS or SR.

Article Snippet: ESCs cultured in monolayer or EB2 cultured in medium supplemented with SR (without LIF) were treated with recombinant WNT11 protein (R&D Systems, Minneapolis, MN, USA) at the final concentration of 250 ng/mL for 48 h and collected for further analyses (referred to as ESC(2) and EB2(2), respectively).

Techniques: Expressing, Control, Cell Culture, Derivative Assay, Incubation, Negative Control, Gene Expression