High glucose increases CTGF mRNA expression (a) and protein production (b, c) in cultured HUVSMCs. Growth-arrested HUVSMCs were stimulated with high glucose (HG, 25 mmol/L) for different durations. (a) Quantitative RT-PCR (Q-PCR) results. Total cellular RNA was isolated from normal glucose (NG, 5.5 mmol/L), high glucose (HG) or mannitol (25 mmol/L) treated HUVSMCs. After reverse transcription, they were subjected to quantitative PCR (Taqman) analysis to determine CTGF mRNA level. Graph is representative of relative CTGF levels in the various conditions. Experiments were performed five times with the similar results (n = 5 in each group). * P < 0.05 vs NG. (b) Representative Western blot (top) and values of total CTGF production (means ± SEM of 3 experiments, bottom). Results of total CTGF protein production were obtained from densitometric analysis and expressed as ratio of CTGF/β-actin. * P < 0.05 vs NG. (c) Immunocytochemical staining of CTGF protein expression in HUVSMCs (top, magnificent of 400×) and integrated optical density (IOD) of the CTGF staining was measured on the images using the Image-Pro Plus software (bottom). Figure shows a representative experiment out of 3 performed experiments. * P < 0.05 vs scrambled-siRNA transfection under normal glucose media condition. # P < 0.05 vs scrambled-siRNA transfection under high glucose media condition. NG: normal glucose; HG: High glucose; scrambled siRNA: scrambled-siRNA plasmid transfection; siRNA: siRNA-CTGF plasmid transfection.
Liu et al. BMC Cell Biology 2007 8:1 doi:10.1186/1471-2121-8-1