bms493  (Tocris)

Journal: bioRxiv

Article Title: Modelling co-development between the somites and neural tube in human Trunk-like Structures (hTLS)

doi: 10.1101/2024.12.16.628661

Figure Lengend Snippet: (a) Schematic diagram of the reciprocal signalling environment of the embryo (left) and hTLS (right). (b) Relative information flow from CellChat analysis, identifying unique signalling interactions between the somite cell cluster and neural clusters (combined Neural progenitor populations 1-3 and Neuronal clusters). Dotted line indicates equal information flow = 0.5. (c) Localised expression of non-canonical Wnt (top) and RA (bottom) signalling components by HCR in 120h hTLS (left), as predicted from CellChat analysis (right). (d-e) Somite-specific ALDH1A2 expression adjacent to the neural tube, in 120h hTLS (d) and E9.5 mouse embryo (e) . (f-g) Projected images of hTLS at 120h following retinoic acid (RA) inhibition at 72h or 96h (BMS493, RA inhibitor). Structures were examined for differences in RA signalling ( ALDH1A2, CYP26A1) and neurogenesis ( DLL1 , NGN1 ; f ) or neural ( SOX2 , PAX6) and posterior tailbud ( WNT3A , FGF8) gene expression (g) . (h) Schematic diagram of proposed reciprocal signalling occurring between somites and the neural tube of hTLSs. Signals from the neural tube bias ALDH1A2 expression in the somites towards the neural tube (left), followed by subsequent RA signalling from somites to neural tube, leading to PAX6 expression biased towards the somitic side (right). RA, retinoic acid; IM, Intermediate mesoderm; LPM, Lateral plate mesoderm. Scale bars, 50 μm; * indicates somites; Dashed line outlines hTLS.

Article Snippet: For RA signalling manipulation of hTLS, retinal (RAL) was replaced with either DMSO or all trans -retinoic acid (100 nM; Sigma Aldrich, R2625); or treated with BMS493 (2.5 µM; Tocris 3509) at 72h during embedding.

Techniques: Expressing, Inhibition

Journal: Development (Cambridge, England)

Article Title: Interplay of SHH, WNT and BMP4 signaling regulates the development of the lamina propria in the murine ureter

doi: 10.1242/dev.204214

Figure Lengend Snippet: Pharmacological analysis of signaling pathways in the early (fetal) development of the lamina propria. (A-D) Embryonic ureters and kidneys were explanted at E12.5 and cultured for 2 days before they were incubated for another 6 days with increasing concentrations of pharmacological inhibitors or activators of SHH signaling [0.1-10 µM cyclopamine (cyclop.), 0.2-10 µM purmorphamine (purmorph.)] (A), WNT signaling (0.1-10 µM IWP-2, 1-10 µM BIO) (B), BMP4 signaling (1-10 µg/ml NOG, 10-250 ng/ml BMP4) (C), and retinoic acid (RA) signaling (0.1-5 µM BMS493, 0.1-5 µM RA) (D). Cultures were then processed for expression of the smooth muscle cell (SMC) marker ACTA2 and the lamina propria (LP) marker ALDH1A2 (left panel), and the total number of cells and the ratio of ACTA2 + ALDH1A2 − SMCs (green), ACTA2 − ALDH1A2 + LP fibrocytes (red), and ACTA2 − ALDH1A2 − ‘undifferentiated’ mesenchymal cells (gray) were determined on proximal ureter sections (right panel). n =5 for each assay. For numbers and statistics (two-tailed Student's t -test, Welch's t -test or Mann–Whitney U -test) see Tables S2A-S9A . * P <0.05; ** P <0.01; *** P <0.001. Data are mean±s.d. c, control.

Article Snippet: For RA signaling manipulation, we used BMS493 (#3509, Tocris; dissolved in DMSO), and RA (#0695, Tocris; dissolved in DMSO).

Techniques: Protein-Protein interactions, Cell Culture, Incubation, Expressing, Marker, Two Tailed Test, MANN-WHITNEY, Control

Journal: Development (Cambridge, England)

Article Title: Interplay of SHH, WNT and BMP4 signaling regulates the development of the lamina propria in the murine ureter

doi: 10.1242/dev.204214

Figure Lengend Snippet: Pharmacological analysis of signaling pathways in the postnatal development of the lamina propria. (A-D) Embryonic ureters were explanted at P0 and cultured for 6 days in the presence of increasing concentrations of pharmacological inhibitors or activators of SHH signaling [0.1-10 µM cyclopamine (cyclop.), 0.2-10 µM purmorphamine (purmorph.)] (A), WNT signaling (1-10 µM IWP-2, 1-20 µM BIO) (B), BMP4 signaling (1-10 µg/ml NOG, 10-250 ng/ml BMP4) (C), and retinoic acid (RA) signaling (0.1-5 µM BMS493, 0.1-5 µM retinoic acid) (D). Cultures were then processed for expression of the smooth muscle cell (SMC) marker ACTA2 and the lamina propria (LP) marker ALDH1A2 (left panel), and the total number of cells and the ratio of ACTA2 + ALDH1A2 − SMCs (green), ACTA2 − ALDH1A2 + LP fibrocytes (red), and ACTA2 − ALDH1A2 − ‘undifferentiated’ mesenchymal cells (gray) were determined on proximal ureter sections (right panel). n =5 for each assay. For numbers and statistics (two-tailed Student's t -test, Welch's t -test or Mann–Whitney U -test) see Tables S10A-S17A . * P <0.05; ** P <0.01; *** P <0.001. Data are mean±s.d. c, control.

Article Snippet: For RA signaling manipulation, we used BMS493 (#3509, Tocris; dissolved in DMSO), and RA (#0695, Tocris; dissolved in DMSO).

Techniques: Protein-Protein interactions, Cell Culture, Expressing, Marker, Two Tailed Test, MANN-WHITNEY, Control

Journal: Development (Cambridge, England)

Article Title: Interplay of SHH, WNT and BMP4 signaling regulates the development of the lamina propria in the murine ureter

doi: 10.1242/dev.204214

Figure Lengend Snippet: Pharmacological analysis of signaling pathway dependency of apoptosis and proliferation in postnatal development of the lamina propria. (A-D) Embryonic ureters were explanted at P0 and cultured for 18 h in the presence of inhibitors of signaling activities [10 µM cyclopamine (cyclop.), 5 µM IWP-2, 10 µg/ml NOG, 1 µM BMS493]. Cultures were then processed for immunofluorescence analysis of the smooth muscle cell marker ACTA2, the urothelial marker CDH1 and TUNEL (A), and for ACTA2, CDH1 and BrdU (B). The BrdU labeling index was determined for ACTA2 − CDH1 − lamina propria fibrocytes (C) and ACTA2 + CDH1 − smooth muscle cells (D). n ≥4 for each assay. For numbers and statistics (two-tailed Student's t -test, Welch's t -test or Mann–Whitney U -test) see Table S20 . * P <0.05; ** P <0.01. Data are mean±s.d. lp, lamina propria; tm, tunica muscularis; ue, ureteric epithelium.

Article Snippet: For RA signaling manipulation, we used BMS493 (#3509, Tocris; dissolved in DMSO), and RA (#0695, Tocris; dissolved in DMSO).

Techniques: Cell Culture, Immunofluorescence, Marker, TUNEL Assay, Labeling, Two Tailed Test, MANN-WHITNEY