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. 2012 May 31;3(5):e316. doi: 10.1038/cddis.2012.57

Figure 2.

Figure 2

Transactivation assay showing LEDGF-dependent activation of Hsp27 in DU145 cells and enhanced cell migration. (A) Representative images showing localization pattern of pEGFP-V (a) or pEGFP-LEDGF (b) in DU145 cells following transfection. Cells were transfected with pEGFP-vector or pEGFP-LEDGF. After 48 h cells were subjected to fluorescence microscopy and photomicrographed. (B) LEDGF overexpression enhanced the transcription of Hsp27. Data showing the real-time PCR analysis of Hsp27 and LEDGF mRNA in cells overexpressing EGFP-LEDGF plasmids as indicated (**P<0.001). (C) Upper panel, diagrammatic representation of Hsp27 promoter containing LEDGF binding elements linked to CAT reporter plasmid. Lower panel, CAT-ELISA performed in DU145 cells after co-transfection with empty CAT vector (black bar) or pCAT-Hsp27 along with varying concentrations of pEGFP-LEDGF (gray bars). (D) DU145 cells overexpressing LEDGF displayed higher migration activity. Wound-healing assay was performed in DU145 cells transfected either with pEGFP-vector or pEGFP-LEDGF. Images are representatives from three independent experiments. Green shows migration of cells transfected with either pEGFP-vector (a, c, e and g) or pEGFP-LEDGF (b, d, f and h). Migration distance was calculated using NIS-Elements BR 3.10 image analyzer software (Nikon, Elgin, IL, USA) and is shown as a histogram (right panel, black bar versus gray bar). Data represent the mean±S.D. from three independent experiments (**P<0.001)