Effects of the Sox17∆50 mutation on SOX17 function in mouse embryonic biliary development
Effects of the Sox17∆50 mutation on SOX17 function in mouse embryonic biliary development
Timed matings between CD1 wild type mice or between Sox17Δ50/Δ50 mice were performed. Wild type and Sox17Δ50/Δ50 embryos were collected separately at E10.5 on ice-cold PBS. Posterior foregut and midgut regions of embryos were dissected by removal of embryonic head, heart, tail, limb buds, posterior and dorsal body trunk. Dissected embryonic tissues were pooled according to genotype and dissociated into single cells by incubating and regular pipetting in Accumax (Millipore Sigma, #A7089) supplemented with DNase I  for 15 minutes at 37C Wild type and Sox17Δ50/Δ50 cell samples were stained separately with Cell Multiplexing Oligo (10x Genomics 3' CellPlex Kit) according to manufacturer recommendations, counted and pooled together at 1:1 ratio in PBS containing 10% FBS on ice. DAPI staining was performed 15 minutes before sorting to enrich live cells at Vanderbilt Flow Cytometry Shared Resource. Library preparation and sequencing was carried out using Chromium Single Cell 3' Reagent Kits v3.1. Libraries were constructed according to the manufacturer's instructions for 10x Genomics Single Cell 3′ Library  kit v3