Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology
Article Title: Alterations in cellular metabolic pathway and epithelial cell maturation induced by MYO5B defects are partially reversible by LPAR5 activation
doi: 10.1152/ajpgi.00091.2024
Figure Lengend Snippet: Lack of effect of Compound-1 on MUC13 localization in MVID model mice. Immunostaining for MUC13 and ACTG1 in jejunal tissues of control, MYO5BΔIEC, and MYO5B(G519R) mice treated with Compound-1 or vehicle. In control jejunum and distal colon, MUC13 (red) is localized to the apical tips of microvilli above actin filaments (green). Both MYO5B loss-of-function mouse tissues demonstrate strong MUC13 immunostaining in cytoplasm of jejunal and colonic epithelial cells. The mislocalization of MUC13 is not remarkably altered by Compound-1 treatments in MYO5BΔIEC or MYO5B(G519R) mice. Scale bars = 20 µm. MYO5B, myosin Vb; MVID, microvillus inclusion disease.
Article Snippet: The primary antibodies against NDUFB8 (1:1,000, Abcam ab192878), HMGCS2 (1:50, Abcam ab137043), NHE3 (1:200, Novus NBP1-82574), SGLT1 (0.5 μg/mL, own) , LAMP1 (1:100, Santa Cruz sc-19992), PHH3 (1:50, Novus NBP3-08511IR, DyLight 750), Ki67 (1:100, Cell Signaling 11882S, Alexa Fluor 488), DCLK1 (1:2,000, Abcam ab202755 Alexa Fluor 647), PCNA (1:50, Santa Cruz sc-56 Alexa Fluor 647), β-catenin/CTNNB1 (12F7) (1:100, Novus NBP1-54467R, DyLight 550), villin (1:50, Santa Cruz sc-58897 Alexa Fluor 488), MUC13 (1:100, Santa Cruz, sc-390115 Alexa Fluor 546), and ACTG1 (1:100, Santa Cruz sc-65638 Alexa Fluor 546 or Alexa Fluor 790) were diluted in Dako antibody diluent with background reducing compound (S3022) and incubated on sections for 1 h at room temperature or overnight at 4°C.
Techniques: Immunostaining, Control