Alexa fluor 488 conjugated donkey anti chicken igg

Immunofluorescence staining and flow cytometry were performed as previously described (18 (link)). Primary antibodies (table S5) were used at the following dilutions: rabbit anti-GFP (Invitrogen, A11122; 1:500), chicken anti-GFP (Abcam, ab13970; 1:1500), anti-αActinin (Sigma-Aldrich, A7811; 1:500), anti-Cx43 (Sigma-Aldrich, C6219; 1:200), anti-Lamp1 (Abcam, ab25245; 1:200), and anti–β-catenin (Abcam, ab16051; 1:200). Secondary antibodies including Alexa Fluor 488–conjugated donkey anti-rabbit IgG, Alexa Fluor 488–conjugated donkey anti-chicken IgG, Alexa Fluor 647–conjugated donkey anti-mouse IgG, cyanine Cy3-conjugated donkey anti-rabbit IgG, and cyanine Cy3-conjugated donkey anti-mouse IgG were all from Jackson ImmunoResearch Inc. Images were captured using EVOS FL Auto Cell Imaging System (Life Technologies). For quantification, 10 to 20 images were randomly taken under ×10 or ×20 magnifications at the same exposure setting and then counted in a double-blinded way. For flow cytometry, iCMs on d10 were harvested by trypsin digestion at 37°C for 5 min. Cells were fixed, permeabilized, probed for αMHC-GFP and cTNT, and then analyzed on a BD Accuri C6 or Cyan flow cytometer. FlowJo software (Tree Star) was used to analyze flow cytometry data.