Immunofluorescent Analysis of Pancreatic Tissues

Pancreatic tissues were fixed for 4 h in 4% paraformaldehyde in PBS and embedded for paraffin sectioning (5 μm). The sections were deparaffinised, rehydrated, and incubated overnight at 4°C with goat antisera against insulin, TCF7L2 antibody (1:60–70, D-4, sc-166699, Santa Cruz, CA, United States), and DAPI (AR1176, Wuhan Boster Company). The sections were subsequently probed with secondary antibodies for 20 min at 37°C (Yang et al., 2012 (link)). Images of the pancreatic tissues were acquired using a fluorescent inverted microscope (Olympus IX71, Japan). For morphometric analysis, the fluorescence intensity of pancreatic sections was quantified using the Image J 1.37c¹.