Titlebook: Hedgehog Signaling; Methods and Protocol Xiaochun Li Book 2022 The Editor(s) (if applicable) and The Author(s), under exclusive license to

協(xié)議

,Expression, Purification, and Structure Determination of Human PTCH1–HH-N Complexes,ng the therapeutic development of cancers. Here, we describe the expression and purification of a nearly full-length functional PTCH1 variant, PTCH1*. With purified PTCH1* protein, two forms of PTCH1*–Sonic Hedgehog (SHH) complexes were assembled, and their structures subsequently determined by cryo-electron microscope (cryo-EM).

事情

Rapid, Direct SMOOTHENED Activity Assays in Live Cells Using cAMP-Based Conformational Sensors,ing changes in SMO conformation within minutes of PTCH1 inactivation by Hh ligands. As a result, it expands the range of experimental perturbations that can be used to dissect PTCH1-SMO communication, enabling a deeper mechanistic understanding of a longstanding mystery in Hh signal transduction.

Cryptic

cortex

Light-Induced Differentiation of Forebrain Organoids by NVOC-SAG,y induce ventral differentiation of human iPS cell-derived forebrain organoids in a light-dependent manner. This article describes preparation of NVOC-SAG, culture of iPS cell-derived forebrain organoids, light irradiation, and downstream analyses.

d-limonene

胡言亂語

Analysis of Dispatched Protein Processing and Sonic Hedgehog Ligand Release,pter, we present a ligand release/retention protocol that allows for the analysis of DISP cleavage, DISP-mediated release of SHH ligand from producing cells, and secretion-dependent signal induction in target cells.

完全

Cell-Based Assays for Smoothened Ubiquitination and Sumoylation,h, which counteracts Smo ubiquitination by recruiting the dub USP8. We describe cell-base assays for Smo ubiquitination and its regulation by Hh and the E3 ligases in .. We also describe assays for Smo sumoylation in both . and mammalian cultured cells.

gentle

Palpate

打火石