Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Spatially restricted dental regeneration drives pufferfish beak development

doi: 10.1073/pnas.1702909114

Figure Lengend Snippet: Gene-expression patterns during dental regeneration morphogenesis and chemical inhibition of Notch signaling through small-molecule treatments. Whole-mount RNA in situ hybridization of the lower jaws of P. baileyi at 57 dpf (A and B and E and F), 53 dpf (H and I), 46 dpf (G), and T. niphobles at 50 dpf (C and D), and P. suvattii at 26 dpf (J and K). (A and B) Images taken from above the lower jaw depict expression of pitx2 (A) and shh (B) in the labial epithelium at the jaw symphysis (white arrowhead). (C–I) Images of a single lower-jaw quadrant, with the expression of pitx2 (C and E), shh (D), lef1 (F), edar (G), notch3 (H), and jagged1b (I) in the developing teeth illustrated through dotted lines (C–G). New tooth units can be seen initially developing (C–F, arrowhead) at the symphysis of the beak (right of image). (J and K) Alizarin red staining of P. suvattii embryos after treatment with 50 μM DAPT during initiation of the second-generation dentition for 72 h, followed by a 2-wk recovery period (K), and control sample treated with 1% DMSO (K). Staining reveals the mineralization of the second-generation tooth (R1, white dotted line), with the control specimens (J) elongating laterally throughout the jaw (n = 5/5). DAPT treatment (25 μM) (K) resulted in the loss of dental elongation, with the mineralized tooth restricted in size at its site of initiation (n = 7/7). (L and M) Schematic representation of the phenotypes observed in the DMSO and DAPT treatments described above. R1–3, replacement tooth generations; S, suture; T1, first tooth generation. (Scale bars: 100 µm in A–F. 50 µm in G–K.)

Article Snippet: DAPT (MedChem Express) stock solution was prepared using DMSO as a solvent.

Techniques: Gene Expression, Inhibition, RNA In Situ Hybridization, Expressing, Staining, Control

Journal: bioRxiv

Article Title: Modulation of host signalling pathways reveal a major role for Wnt signalling in the maturation of Plasmodium falciparum liver schizonts

doi: 10.1101/2024.05.07.592925

Figure Lengend Snippet: A) Dot plots representing pathways that are upregulated in hepatocytes cultured in B27 for 7 days post plating. Heatmaps representing normalized expression of genes from (B)Wnt, (C)TGFβ, (D)Notch, (E)BMP and Adenylate cyclase signalling pathways deregulated in cultured vs freshly isolated hepatocytes. The data is scaled and representative of 2 biological replicates of RNA sequencing. The effect of specific host pathway inhibitors (IWP2, SB431542, DAPT, LDN193189 and Forskolin) on respectively schizont numbers (F; each dot is the average count of two technical replicates), size of schizonts (G; each dot is the median of at least 100 schizonts), total DAPI per schizont (H; each dot is the median of at least 100 schizonts), and total PfMSP1 per schizont (I; each dot is the median of at least 100 schizonts) on day 5 p.i.. The grey dotted line (per graphs F-I) shows the median measurements of schizonts grown in B27 control (from four independent experiments). The p-values from a Dunnett’s multiple comparisons test are shown. J) Representative confocal images (from four independent experiments) showing schizonts grown in B27 alone and B27 supplemented with Forskolin, SB431542, IWP2, DAPT and LDN193189 on day 5 p.i. stained with PfMSP1, PfHSP70 and DAPI. Scale bar is 25 microns.

Article Snippet: The following day, it was changed to the following medium conditions: B27 or B27 supplemented with 20μM of Forskolin (Enzo Life Sciences: BML-CN100-0010), or 10μM of SB431542 (Tocris: 1614), or 0.5μM of IWP2 (Tocris: 3533), or 5μM of DAPT (Tocris: 2634) or 0.1μM of LDN193189 (Tocris: 6053) or the combination of all the compound inhibitors (i.e. 5C).

Techniques: Cell Culture, Expressing, Isolation, RNA Sequencing, Control, Staining