Bx41 microscope

Tissue biopsies were directly embedded in the OCT compound or fixed with Carnoy’s fluid. Paraffin-embedded tissues were used for hematoxylin/eosin (H&E) staining, and frozen sections were fixed in 4% PFA for 20 min to immunofluorescence staining. For Gram staining, tissue is fixed with Carnoy’s fluid and embedded in OCT compound, sectioned by 4um and stained with Gram stain (Remel, Lenexa, KS). For Periodic acid-Schiff(PAS) staining, tissue is fixed with Carnoy’s fluid and embedded in Paraffin, and stained with PAS. For TUNEL staining, TUNEL Andy Fluo 594 Apoptosis Detection Kit (#A051, abpbio) is used. For IHC, fixed and permeabilized frozen tissue sections were blocked with Image-iT FX reagent (Invitrogen) before incubating with HABP (#385911, EMD Millipore), Muc2 (#PIMA512345, Fisher) or Reg3g (#PA5-50450, Thermo Fisher) in 1:100 dilution followed by appropriate 488- or 568-coupled secondary antibodies. Nuclei were counterstained with DAPI. All images were taken with an Olympus BX41 microscope (widefield) or Zeiss LSM510 confocal microscope as indicated.

NHEKs were grown on 8-well chamber slides (Thermofisher Scientific). After indicated treatments, cells were fixed in 4% paraformaldehyde (PFA) (Thermofisher Scientific) for 10 minutes and blocked with 3% bovine serum albumin, 0.2 M Glycine (Sigma) with or without 1 mg/mL saponin (Sigma) prior to incubating with primary antibodies followed by appropriate 488- or 568-coupled secondary antibodies, or 488-streptavidin conjugate. Nuclei were counterstained with DAPI. All images were taken with an Olympus BX41 microscope (widefield), Zeiss LSM510 confocal microscope, or Nikon A1R Confocal STORM microscope as indicated.