Tween 20 detergent

All
aptamers used in this work (Table 6) were acquired from
Integrated DNA Technologies.
Phosphate-buffered saline (PBS) was made up
in milliQ water as
a solution of 10 mM Na₂PO₄ (ECP Labchem), 1.8
mM KH₂PO₄ (ECP Labchem), 137 mM NaCl (Sigma
Aldrich), and 2.7 mM KCl (Sigma Aldrich). To this solution, 0.005%
by volume Tween-20 detergent (ThermoFisher Scientific) was added to
form the SPR buffer (PBS-T). Tris buffer was made up as a 1 M solution
of Tris base (ThermoFisher Scientific) in milliQ and adjusted to pH
8 using hydrochloric acid.
Analytical grade silver nitrate (99.8%,
Aldrich), trisodium citrate
(99.0%, BDH Chemicals), hydrogen peroxide (30% w/w, ThermoFisher Scientific),
potassium bromide (99.0%, Ajax Finechem), and sodium borohydride (98.0%,
Aldrich) were used in nanoparticle preparation.

Total protein from the testes samples and in vitro cultured Sertoli cells was isolated using a Proprep kit (17081; iNtRON) according to the manufacturer’s instructions. Fifty micrograms of total protein from each sample were separated using 4–20% gradient SDS-polyacrylamide gel electrophoresis (Bio-Rad Laboratories) and transferred to polyvinylidene fluoride membranes. The membranes were blocked with 5% non-fat milk and incubated for 1 h at room temperature with the primary antibodies for each protein: β-actin and clusterins 2, 3, 5, 11, 18. After washing three times with tris-buffered saline with Tween™ 20 detergent (Thermo Fisher Scientific, Inc.), after which the appropriate secondary antibodies were added. The membranes were then incubated for 2 h at room temperature, after which protein expression was confirmed using enhanced chemiluminescence (32106; Thermo Fisher Scientific, Inc.) with HyBlot CL^® Autoradiography Film (No. E3018; Denville Scientific, Metuchen, NJ, USA). The antibodies used for Western blot analysis of claudins and other marker proteins are listed in Table 1.