Fig. 1.
Ectopic expression of Survivin in different cell lines. a Scheme of the Survivin (top) and control (bottom) constructs including the coding sequence for Survivin fused to c-myc and HA tags. T2A: self-cleaving 2A peptide; puroR: puromycin resistance. b Western blot analysis demonstrating ectopic Survivin in U251-MG, SVGp12, U87-MG, U87-MGshp53, HCT116 and HCT116p53−/− cells. As control empty vector-transduced cells were used. Cells were either transduced with lentiviral pHATrick-Survivin-EGFP (Surv) or empty vectors (C) and lysed 72 h after transduction. Ectopic Survivin (24 kDa) (black arrows) and endogenous Survivin (16.5 kDa) (asterisks) bands are indicated. The membranes were re-probed with antibodies specific for α-actin (42 kDa) to confirm equal loading. c Indirect immunofluorescence analysis using specific anti-Survivin antibodies revealed typical immunosignals in for endogenous Survivin and for Survivin-transduced cells at the cleavage furrow and midbodies (arrows). Note the damaged lagging chromosome which seems broken after cytokinesis (white arrowheads). From left to right: DNA (DAPI), Survivin (FITC), membrane (WGA-TexasRed) and merge