Cd42 fitc

Blood samples were obtained after 12 hours of fasting and the analyses were
performed at the central laboratory of our university. All athletes were
allowed to maintain their daily exercises program even on the day before
blood sample collection. The athletes had very similar exercise training
programs, corresponding to two long-distance running sessions every day, 15
km in the morning and 10 km in the afternoon, and intensive training
(100-1,000 meter shots, repeated many times)twice a week, on Tuesday and
Thursday mornings. All blood samples were collected on Thursdays, before
exercise.
Measurements of EPCs and MPs were performed as previously reported, using
fresh blood samples in EDTA containing tubes.^{12 (link)-15 (link)}For determination of EPCs, a minimum of 500,000 events was acquired by
flow-cytometry (FACSCalibur, BD Biosciences, USA). Fluorescently labeled
mouse anti-human antibodies were used for EPCs (CD34 FITC, BD Biosciences,
USA; CD133 APC, Miltenyi Biotec, USA; KDR PE, R&D Systems, USA), PMPs
(CD42 FITC and CD31 PE, BD Biosciences, USA) and EMPs (CD51 FITC, BD
Biosciences). Disposable containers (BD Biosciences) were used to quantify
the number of microparticles per microliter of
platelet-poor plasma (PPP).

TRIzol was purchased from Sigma (St Louis, Missouri, USA). TMR red-labeled In-Situ Cell Death Detection reagent and sTREM-1, the sSirt6 enzyme-linked immune sorbent assay (ELISA) kit were purchased from Roche Applied Science (Indianapolis, USA). Monoclonal antibodies against CD62-PE, CD31-PE, CD42-FITC, and IgG-PE were purchased from BD (Shanghai, China), and monoclonal rabbit antibodies against Sirt6, TREM-1, LC3, BECN1, LAMP2, p62, Caspase-1, and GAPDH were purchased from Cell Signaling (Denver, Colorado, USA). Fetal bovine serum (FBS) and Lipofectamine^® 2000 Transfection Reagent were purchased from Invitrogen (Carlsbad, CA, USA). The cDNA Synthesis Kit and Premix Ex Taq SYBR Green PCR Kit were purchased from Takara (Shiga, Japan). The adv-Sirt6, LV-Sirt6 shRNA, adv-TREM-1, LV-TREM-1 shRNA, LV-LC3 shRNA, and GFR, GRP double labeled adv-LC3 were purchased from HanBio. (Shanghai, China). Other unmentioned reagents were purchased from Shenggong Bio. (Shanghai, China).

Glucose and TRIzol were purchased from Sigma (St Louis, Missouri, USA). Monoclonal antibodies against CD62-PE, CD31-PE, CD42-FITC and IgG-PE were obtained from BD (Shanghai, China), and the monoclonal rabbit antibodies against Sirt6, p-eNOS, eNOS and β-actin were obtained from Cell Signaling (Denver, Colorado, USA). The fetal bovine serum (FBS), DMEM culture medium with or without glucose, Lipofectamine® 2000 Transfection Reagent and ECMatirm™ solution were purchased from Invitrogen (Carlsbad, CA, USA). The cDNA Synthesis Kit and Premix Ex Taq SYBR Green PCR Kit were purchased from Takara (Shiga, Japan). The adv-Sirt6, Sirt6 siRNA, CMV-Sirt6 shRNA, Sirt1 siRNA and Drosha siRNA were obtained from HanBio. (Shanghai, China). Other unmentioned reagents were purchased from Shenggong Bio. (Shanghai, China).