Figure 2

Inhibition of the soluble uPAR by knockdown of HSP70 and/or MRJ in HCT116 cells. (A) Effects of siRNAs targeting uPAR, HSP70 and MRJ by Western Blot analysis. The cells were transfected with psiuPAR, or psiHSP70, or psiMRJ or psiSC as shown in the figures. The levels of uPAR, HSP70 and MRJ proteins were normalized to GAPDH levels in psiSC-transfected cells. (B) Knockdown of HSP70/MRJ reduced the protein levels of suPAR and HSP70. The cells were transfected with psiSC, or psiMRJ, or psiHSP70, or psiMRJ plus psiHSP70 for 48 hours and processed and then probed with anti-uPAR antibody. The blots were re-probed with GAPDH to confirm equal amounts of protein loading. Protein band intensities were quantified by densitometry analysis using Image J software. Bars indicate the values expressed as the mean ± standard deviation. Statistical evaluation showed that the changes were significant as shown by *P < 0.05. Data shown are representative of three independent experiments.