Male reproductive system ageing is closely associated with deficiency in testosterone production due to loss of functional Leydig cells, which are differentiated from stem Leydig cells (SLCs). However, the relationship between SLC differentiation and age
were dried entirely in a vacuum centrifuge. For Western blot analysis, the dried EV samples were lysed directly in LDS buffer (lower volumes of urine (~0.5–2 mL) were used for Western blot experiments). EV isolation by differential ultracentrifugation (UC) The EV isolation by UC was per...
were dried entirely in a vacuum centrifuge. For Western blot analysis, the dried EV samples were lysed directly in LDS buffer (lower volumes of urine (~0.5–2 mL) were used for Western blot experiments). EV isolation by differential ultracentrifugation (UC) The EV isolation by UC was per...
Cells were incubated in buffer A (10 mM Tris, 1 mM KCl, 1.5 mM MgCl2) for 10 min on ice then dounce homogenized with 10 strokes. The mixture was centrifuged, and the pellet was resuspended in buffer B (50 mM Tris-HCl, 10 mM EDTA, 1% SDS) and kept for 10 min on ice. DNA was...
All lyophilized samples were redissolved in 0.1% formic acid (buffer A) at a concentration of 5 mg/mL before MS analysis. All peptide mixtures were analyzed on a reverse phase C18 capillary LC column from Michrom Bioresources (100 μm x 150 mm, 3 μm, 0.5 μL/min). The elution grad...
After stimulation with the indicated ligand for 5 min, the cells were lysed in 80 μL lysis buffer (Beyotime, China) containing protease inhibitors (Roche Applied Science) at 4 °C for 30 min on a rocker and then scraped. Equal amounts of total cell lysate were size fractionated using ...
The cells were then scraped into 1 ml of 100 mm sodium bicarbonate buffer, pH 11.5, and homogenized by 5 strokes in a 2-ml Dounce homogenizer. After 30 min incubation at 4 °C, the homogenates were centrifuged at 100,000 rpm at 4 °C in a TLA 100.2 rotor (Beckman). The pellets ...
Buffer BuildingMinds DigitalTwin Core BulkSMS Business Assist Businessmap Buy Me A Coffee (Independent Publisher) Byword (Independent Publisher) Calculate Working Day Calendar Pro Calendarific (Independent Publisher) Calendly Calendly (legacy) Campfire CandidateZip Resume/Job Parser Capsule CRM Captisa Form...
Buffer BuildingMinds DigitalTwin Core BulkSMS Business Assist Businessmap Buy Me A Coffee (Independent Publisher) Byword (Independent Publisher) Calculate Working Day Calendar Pro Calendarific (Independent Publisher) Calendly Calendly (legacy) Campfire CandidateZip Resume/Job Parser Capsule CRM Captisa Form...
Cells were lysed in cell lysis buffer containing 50 mM Tris–HCl, pH 8.0, 150 mM NaCl, 1% IGEPAL CA-630, and 100 mM phe- nylmethylsulfonyl fluoride and 5% protease inhibitor cocktail (Sigma-Aldrich). Lysis took place on ice for 30 min, followed by centrifugation at 14,000 g, 4 °C...