notch signaling pathway inhibitor dapt  (Bioscientifica Ltd)

Journal: Biology Open

Article Title: Apoptosis is a generator of Wnt-dependent regeneration and homeostatic cell renewal in the ascidian Ciona

doi: 10.1242/bio.058526

Figure Lengend Snippet: Pharmacological screen and siRNA effects on oral siphon regeneration. (A) Bar graphs showing the effects of signaling system inhibitors on percent oral siphon regeneration. Percent oral siphon regeneration was determined with respect to controls treated with DMSO. The inhibitors and affected signaling systems are shown at the top of the bars. The numbers of animals assayed are shown at the bottom of the bars. Black asterisks indicate significant differences at P <0.001 between the control and inhibitor treated animals. Red asterisk indicates significant difference at P <0.01. Statistics by χ 2 test and post-hoc Fisher's exact test with Bonferroni correction. (B–J) Effects of siRNA on oral siphon regeneration. (B,C,E,H) Normal regeneration after treatment with (B) ma3 siRNA, (C) scrambled delta1 siRNA, (E) scrambled ß-catenin siRNA, or (H) scrambled wnt3 siRNA. (D,F,G,I) Suppression of regeneration after treatment with (D) delta1 siRNA, (F) ß-catenin siRNA, (G) ß-catenin siRNA and Wnt3a, or (I) wnt3 siRNA. (J) Rescue of regeneration after treatment with wnt3 siRNA and Wnt3a. Arrows: oral siphon pigmented organs. (K) RT-PCR showing delta1 , ß-catenin , and wtn3 mRNA expression at 6 days post-amputation without corresponding siRNA (-siRNA) and with corresponding siRNA (+siRNA) and notch expression without wnt3 siRNA (−siRNA) and with wnt3 siRNA (+ wnt3 siRNA). (L) Bar graphs showing the effects of siRNA on percent oral siphon regeneration and rescue by exogenous Wnt3a. The number of animals used in each experiment is shown at the bottom of the bars. Black asterisks indicate significant differences at P <0.001 between the respective scrambled controls and the corresponding siRNA treated animals. Red asterisk indicates significant difference at P <0.01. Statistics by χ 2 test and post-hoc Fisher's exact test with Bonferroni correction. Each experiment was replicated at least two times.

Article Snippet: The FGF signaling pathway inhibitor was SU5402 (Tocris Bioscience, Bristol, UK) (5 μM) , the Hedgehog signaling pathway inhibitors were cyclopamine (Tocris Bioscience) (20 μM) ( ) and Sant-1 (Tocris Bioscience) (15 μM) , the BMP signaling pathway inhibitor was dorsomorphin (Tocris Bioscience) (5 μM) , the Notch signaling pathway inhibitors were DAPT (Tocris Bioscience) (12 μM) ( ) and Compound E (Abcam, Cambridge, MA, USA) (1 μM) , and the Wnt pathway signaling inhibitors were FH535 (Santa Cruz Biotechnology Inc., Dallas, TX, USA) (1.2 μM) ( ) and IWR-1-Endo (Santa Cruz Biotechnology) (2.5 μM) ( ).

Techniques: Control, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: TNFSF15 facilitates human umbilical cord blood haematopoietic stem cell expansion by activating Notch signal pathway

doi: 10.1111/jcmm.15626

Figure Lengend Snippet: TNFSF15 facilitates human umbilical cord blood haematopoietic stem cell expansion by Notch signal pathway. A, The representative plots of flow cytometry analysis and (B) the statistics of CD34 + cell apoptosis rates in the presence or absence of TNFSF15 (2 µg/mL). The experiment was repeated for three times (n = 3). C, The representative plots of flow cytometry analysis and (D) the statistics of CD34 + cell cycle distribution of G0/G1, S and G2‐M phase in the presence or absence of TNFSF15 (2 µg/mL) for 7 d in expansion medium (n = 3). The experiment was repeated for three times. E, The percentage of G0 phase was detected in the presence or absence of TNFSF15 (2 µg/mL) for 7 d in expansion medium by Ki67/PI double staining assay (n = 3). F, The gene expression of c‐myc in CD34 + CD38 − CD90 + CD45RA − CD49f + cells after the treatment of TNFSF15 (2 µg/mL) for 7 d in expansion medium (n = 3). G, The gene expression of hes1 in CD34 + CD38 − CD90 + CD45RA − CD49f + cells after the treatment of TNFSF15 (2 µg/mL) for 7 d in expansion medium (n = 3). H, Western blotting analysis of Notch signal pathway proteins including c‐myc, hes1, Notch1 and NCID in CD34 + cells treated with TNFSF15 at indicated concentrations for 7 d in expansion medium. I, The absolute number of CD34 + CD49f + cells after cultured with combination of TNFSF15 (2 µg/mL) and DAPT (0.5 µmol/L) for 7 d in expansion medium (n = 3)

Article Snippet: The small compounds SR1, UM171 and Notch signal pathway inhibitor DAPT (GSI‐IX) were purchased in Selleck.

Techniques: Flow Cytometry, Double Staining, Gene Expression, Western Blot, Cell Culture