Goat anti dll4

The following primary antibodies were used: rabbit anti-FSCN1 (Abcam, ab126772; IF 1:200, WB 1:1000, IP 1:100), mouse anti-GAPDH (Abcam, ab8245; WB 1:1000), mouse anti-F-actin (Abcam, ab205; WB 1:1000), rabbit anti-YAP (Cell Signaling Technology, 14074; IF 1:200, WB 1:1000), rabbit anti-p-YAP (Ser127) (Cell Signaling Technology, 4911; WB 1:1000), mouse anti-CDC42 (BD Biosciences, 610929; WB 1:500), rabbit anti-VEGFR2 (Cell Signaling Technology, 2479; WB 1:1000), rabbit anti-p-VEGFR2 (Tyr1175) (Cell Signaling Technology, 2478; WB 1:1000), goat anti-DLL4 (R&D Systems, AF1389; IF 1:200), and mouse anti-β‐actin (Millipore, A5441; WB 1:1000). The following secondary antibodies were used: goat anti-mouse HRP-conjugated antibodies (Beyotime, A0216; 1:1000) and goat anti-rabbit HRP-conjugated antibodies (Beyotime, A0208; 1:1000) were applied for WB, Alexa Fluor™ Plus 488 Anti-mouse IgG (H + L) (Abcam, ab150077; IF 1:400), and DyLight 405-AffiniPure Donkey Anti-goat IgG (H + L) (Jackson ImmunoResearch,705–475-147,1:400) was applied for immunofluorescence.

Kaiso^Tg intestinal tissues were formalin-fixed and paraffin embedded as previously described [19 (link)]. Periodic acid-Schiff (PAS) staining was performed by the John Mayberry Histology Facility at McMaster University. Immunohistochemistry (IHC) analysis of all other protein targets was performed as previously described [19 (link)], with the following modifications: antigen retrieval for chromogranin A was accomplished by heating tissues in 10 mM sodium citrate, pH 6.0 for 10 min at sub-boiling temperature; retrieval for NICD, Dll-1, Dll-4, Hes1, and Hes5 was accomplished by heating tissues at sub-boiling temperature for 15 min in TE-Tween (Tris EDTA, 0.05% Tween), pH 9.0; and retrieval for lysozyme was performed with 200 μg/mL proteinase K, 50 mM Tris pH 7.4 at RT for 5 min. Tissues were incubated with the following primary antibodies overnight at 4 °C at the indicated dilutions: rabbit anti-lysozyme (Thermo Scientific cat. #PA1–29680; 1:50); rabbit anti-chromogranin A (Abcam cat. #ab15160; 1:500); rabbit anti-Cleaved Notch 1 Val-1744 (Cell Signaling Technology cat. #4147; 1:75); rabbit anti-Hes1 (Cell Signaling Technology cat. #11988S; 1:80); rabbit anti-Hes5 (Abcam cat. #ab65077; 1:125); rabbit anti-Dll-1 (Abcam cat. #ab84620; 1:100); and goat anti-Dll-4 (R&D Systems cat. #AF1389).