Glucose Oxidase Induces Cellular Senescence in Immortal Renal Cells through ILK by Downregulating Klotho Gene Expression

Figure 3

ILK mediates cellular senescence induced by GOx. (a) MCT cells were transfected with a plasmid coding for wild-type ILK protein (WT-ILK) at several doses (0, 1, or 2 μg). ILK and p16 proteins were evaluated by western blot. (b, c) MCT were transfected with the specific siRNA for ILK (siILK) or unspecific scramble siRNA (scRNA) as control and then treated with 2.5 mU/mL of GOx for 48 h. In those conditions, mRNA ILK was evaluated by RT and real time PCR (b) and ILK and p16 protein expression was evaluated by western blot (c). A representative blot is shown in each case. Bar graphs represent the densitometric analysis of the bands. The results are expressed as densitometric units and are the mean ± s.e.m. of the mean from five different experiments. versus control; versus scRNA.

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