As a pilot test to determine the most efficient treatment time and concentration of Epo, levels of connective tissue growth factor (CTGF) and transforming growth factor-β (TGF-β) were measured by real-time PCR, after treatment with Epo on MGCs cultured in high glucose.  Matrix metalloproteinase 9 (MMP9) mRNA level was detected by real-time PCR. 
real-time PCR
Protein levels of CTGF and cytoskeletal proteins like α-SMA and fibronectin were measured by enzyme-linked immunosorbent assay (ELISA) and Western blot respectively. 
ELISA
Protein levels of CTGF and cytoskeletal proteins like α-SMA and fibronectin were measured by enzyme-linked immunosorbent assay (ELISA) and Western blot respectively. 
Western blot
Wound-healing assay was applied to evaluate the migratory ability of MGCs, and actin-tracker green was used to draw the structure of F-actin in MGCs.



actin-tracker green