Immunohistochemical Analysis of Exoc5 Mice

Immunohistochemical and fluorescence stains were performed on 5 um, paraffin-embedded sections from E13.5, E15.5, P0, and adult wildtype and conditional Exoc5 mice as previously described^{11 (link)}. Antibodies and their dilutions include: acetylated tubulin (Sigma, #T6793, 1:500), gamma tubulin (Abcam, #T6793, 1:1000), RUNX2 (Abcam, #ab192256, 1:250), ERK1/2 (Cell Signaling, #4695, 1:50), phospho-ERK1/2 (Cell Signaling, #4370, 1:40), cleaved NOTCH1 (Cell Signaling, #4147, 1:200), Hoescht (Life Technologies, #H3569, 1:10,000), and Life Technologies Alexa Fluors: 488 (#A-11029, #A-11034) 568 (#A-11004, #A-11036) and Cy5 (#A-10524) at 1:100. Slides were coverslipped using Invitrogen SlowFade Gold Antifade Reagent (#S36936). Images were captured using: Leica TCS SP5 AOBS Confocal Microscope System and LAS AF v2.6.3 Build 8173 Acquisition and Analysis Software, Zeiss Axioscope M2, or Olympus BH-2 brightfield microscope.

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Fulmer D., Toomer K., Guo L., Moore K., Glover J., Moore R., Stairley R., Lobo G., Zuo X., Dang Y., Su Y., Fogelgren B., Gerard P., Chung D., Heydarpour M., Mukherjee R., Body S.C., Norris R.A, & Lipschutz J.H. (2019). Defects in the exocyst-cilia machinery cause bicuspid aortic valve disease and aortic stenosis. Circulation, 140(16), 1331-1341.

Publication 2019

acetylated tubulin gamma tubulin RUNX2 ERK1/2 phospho-ERK1/2 cleaved NOTCH1 SlowFade Gold Antifade Reagent TCS SP5 AOBS Confocal Microscope System Axioscope M2 BH-2 brightfield microscope

Adult Antibodies Confocal microscope Dilutions Embedded paraffin Erk1 Fluorescence Gamma tubulin Gold Mice Microscope Runx2 Stains Tubulin

Corresponding Organization :

Other organizations : Medical University of South Carolina, University of Hawaiʻi at Mānoa, Clemson University, Brigham and Women's Hospital, Harvard University, Ralph H. Johnson VA Medical Center, Beth Israel Deaconess Medical Center

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Variable analysis

independent variables

Genotype (wildtype and conditional Exoc5 mice)

dependent variables

Acetylated tubulin
Gamma tubulin
RUNX2
ERK1/2
Phospho-ERK1/2
Cleaved NOTCH1

control variables

Developmental stages (E13.5, E15.5, P0, and adult)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

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