Figure 4.

Up-regulation of ILK protein level in response to high concentration of glucose. Rat mesangial cells were cultured in RPMI 1640 medium containing a normal concentration (5.6 mmol/L) (lane 1) or a high concentration (30 mmol/L) (lane 2) of glucose for 21 days. ILK and actin in cell lysates (0.25 mg/ml) were detected by immunoblotting (20 μl lysates/lane) with monoclonal anti-ILK antibody 65.1 (A) or anti-actin antibody mAb1501 (B) as described in Materials and Methods. Note that the amount of ILK (59 kd) was significantly increased in mesangial cells exposed to high glucose concentration. The amount of a higher molecular mass band (80 kd) that was recognized by the monoclonal anti-ILK antibody, which likely represents a sodium dodecyl sulfate-resistant ILK-containing protein complex or an ILK variant, 41 was also increased in response to high glucose.