Periodic acid schiff stain kit

Adenosine and cordycepin were purchased from Sigma Chemical Co. (St. Louis, MO, USA). The hematoxylin and eosin (HE) stain kit (CATA: ab245880) and periodic acid Schiff (PAS) stain kit (CATA: ab150680) were purchased from Abcam (Cambridge, MA, USA).

BALB/c female mice of 5–7 weeks old were procured from the institute animal house and were maintained in individually ventilated cages under standard ad libitum conditions. Oligonucleotides used in RT-PCR were procured either from Eurofins scientific or Integrated DNA Technologies. Periodic Acid Schiff (PAS) Stain Kit from Abcam, Cambridge, MA, USA, was obtained. C. albicans (SC5314/WT), HLC52 (defective in efg1), HLC54 (defective in both cph1 and efg1), and JKC19 (defective in cph1) strains were cultured in liquid YPD growth media (1% yeast extract, 2% peptone, and 2% glucose in distilled water), whereas the two bacterial strains, S. aureus (ATCC43300) and E. coli (MG1655), were cultured as a monoculture in BHI (7.7 g of calf brain, 9.8 g of beef heart, 10 g of protease peptone, 2 g of dextrose, 5 g of sodium chloride, and 2.5 g of disodium phosphate per liter) and LB (10 g of tryptone, 5 g of yeast extract, and 10 g of NaCl per liter) nutrient media, respectively. For coculture experiments, YPD was used.

Mice testes and cauda epididymis were isolated immediately after euthanasia, fixed in Bouin's solution (Sigma, #HT10132-1L) for 24 h, and then embedded in paraffin. Five-micron sections were prepared and mounted on glass slides. After deparaffinization, slides were stained with hematoxylin and eosin (HE) for histological analysis. TUNEL staining was performed with TUNEL kit (keyGEN BioTECH, #KGA7072) following the manufacturer's instructions. PAS staining was performed with Periodic Acid Schiff (PAS) Stain Kit (Abcam, ab150680). Stages of seminiferous epithelium cycle and spermatid development were determined as previously described (36 (link)). For chromosome spread, testicular tubules were pretreated by hypotonic buffer (30 mM Tris, 50 mM sucrose, 17 mM trisodium citrate dihydrate, 5 mM EDTA, 0.5 mM DTT and 0.5 mM phenylmethylsulphonyl fluoride (PMSF), pH 8.2) for 30 min. Short fragments of testicular tubules were suspended in 0.1 M sucrose and dispersed to single cells, then spread to a thin cell layer on glass slides, and treated with 1% (w/v) paraformaldehyde solution containing 0.15% Triton X-100 (37 (link)).

Colon tissues were rolled and fixed in 10% neutral formalin, and 3 μm paraffin sections were stained with hematoxylin and eosin (H&E). The degree of inflammation was measured by a pathologist using i-Solution Lite (IMT i-Solution Inc., Daejeon, Korea). The pathological grade is expressed as a percentage^{32 (link)}: 0, normal; g1, intestinal gland loss ≤ 25% of the LP mucosae and slightly increased inflammatory cell infiltration of the LP; g2, intestinal gland loss ≥ 25% or ≤ 50% and significantly increased inflammatory cell infiltration; and g3, intestinal gland loss ≥ 50% with severe erosion of inflammatory cells. To visualize mucin and goblet cells, colon sections were stained with a Periodic acid Schiff (PAS) stain kit (Abcam, Cambridge, UK) according to the manufacturer’s instructions. The number of goblet cells per crypt was determined using a Nikon Eclipse Ts2 (Nikon, Minato, Tokyo, Japan).