Spraque-Dawley rats, weighing 180 250 gr (Harlan laboratories, Jerusalem, Israel)were anesthetized by intraperitoneal injection of the combination of Ketamine, 80 mg/kg and Xylazine, 10 mg/kg.
GR and RM performed laboratory procedures, FACS-sorting and mammosphere cultures.
Work in our laboratory identified GR amongst factors that were up-regulated during chondrocyte maturation [ 50] Thus, to comprehensively understand the transcriptional effects of pharmacological GC doses in growth plate, we completed a genomic screen of gene expression changes in chondrocytes derived from E15.5 day old mouse embryos.
The protein was probed with polyclonal anti-GR antibody made by our laboratory (1 1,000), monoclonal anti-ER antibody (1 750; Immunotech, Marseille, France), polyclonal anti-GILZ antibody (1 500; Santa Cruz Biotechnology, Santa Cruz, CA), or monoclonal anti-β-actin antibody (1 10,000; Millipore, Temecula, CA).
Naturally, occurring glucocorticoids (cortisol and hydrocortisone) dissociate within 15 min from GR, and any remaining unbound fraction would be washed away in the laboratory process.
The stool specimens (0.5 1.5 gr) were collected in labeled plastic vials without preservatives and transported to the laboratory within four hours after collection.
MG and GR conceived the study, supervised the data analysis and drafted the manuscript; PCF and CC carried out all the laboratory and serological assays; ADC and GP enrolled the study participants; SB contributed to performing the statistical analysis.
GR carried out renal scintigraphy 7. MR magnetic resonance performance 8. GV collaboration in renal scintigraphy 9. DS clinical laboratory work 10.
(in part) and by grants from the Ministry of Research (France); University Paris Diderot (France); National Institute for Blood Transfusion (INTS, France); National Institute for Health and Medical Research INSERMM, France); 'Investissements d'avenir', Laboratories of Excellence GR-Ex (to A.G.deB).
