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1. Generation and Analysis of Mosaic Spinal Cord Organoids Derived from Mouse Embryonic Stem Cells
Carina Jägers and Henk Roelink
2. Evaluating Hedgehog Acyltransferase Activity and Inhibition Using the Acylation-Coupled Lipophilic Induction of Polarization (Acyl-cLIP) Assay
Sebastian A. Andrei, Thomas Lanyon-Hogg, and Edward W. Tate
3. Analysis of Protein Cholesterylation by Biorthogonal Labeling
Ao Hu, Meng Zhou, and Bao-Liang Song
4. An In Situ Fluorescence Assay for Cholesterol Transporter Activity of the Patched
Wonhwa Cho, Arthur Ralko, and Ashutosh Sharma
5. Imaging of the Ciliary Cholesterol Underling the Sonic Hedgehog Signal Transduction
Tatsuo Miyamoto, Kosuke Hosoba, Silvia Natsuko Akutsu, and Shinya Matsuura
6. Studying Hedgehog Signaling during Mouse Neural Tube Development
Sandii Constable, Kevin White, Bandarigoda Nipunika Somatilaka, and Saikat Mukhopadhyay
7. Measuring and Manipulating Membrane Cholesterol for the Study of Hedgehog Signaling
Maia Kinnebrew, Kristen A. Johnson, Arun Radhakrishnan, and Rajat Rohatgi
8. The Immunofluorescence-Based Detection of Hedgehog Pathway Components in Primary Cilia of Cultured Cells
Shu Yang and Xiaoyan Zheng
9. Analysis of Dispatched Protein Processing and Sonic Hedgehog Ligand Release
Elizabeth Cleverdon, Daniel P. Stewart, and Stacey K. Ogden
10. Expression, Purification, and Structure Determination of Human PTCH1–HH-N Complexes
Xiaofeng Qi, Philip Schmiege, Leticia Esparza, and Xiaochun Li
11. Characterization of Smoothened Phosphorylation and Activation
Jianhang Jia, Yuhong Han, Yong Suk Cho, and Jin Jiang
12. Cell-Based Assays for Smoothened Ubiquitination and Sumoylation
Yuhong Han and Jin Jiang
13. Biochemical Assays to Directly Assess Smoothened Activation by a Conformationally Sensitive Nanobody
Ishan Deshpande and Aashish Manglik
14. Studies of SMOOTHENED Activation in Cell-Free and Reconstituted Systems
Isaac B. Nelson and Benjamin R. Myers
15. Rapid, Direct SMOOTHENED Activity Assays in Live Cells Using cAMP-Based Conformational Sensors
Madison F. Walker and Benjamin R. Myers
16. Light-Induced Differentiation of Forebrain Organoids by NVOC-SAG
Ryuji Misawa and Yoshiho Ikeuchi
17. NanoBRET and NanoBiT/BRET-Based Ligand Binding Assays Permit Quantitative Assessment of Small Molecule Ligand Binding to Smoothened
Pawel Kozielewicz and Gunnar Schulte
18. Measuring Smoothened (SMO)-Mediated Activation of the Gi Protein
Heng Liu and Cheng Zhang
19. Ci/Gli Phosphorylation by the Fused/Ulk Family Kinases
Yuhong Han and Jin Jiang
20. A Method for Orthotopic Transplantation of Lung Cancer in Mice
Alexandra N. Wilson, Baozhi Chen, Xin Liu, Jonathan M. Kurie, and James Kim
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