Tetracycline-inducible protein expression in pancreatic cancer cells: Effects of CapG overexpression

Figure 7 Subcellular localization of CapG in stable CapG inducible clones by immunohistochemistry (A) and subcellular fractionation (B). A: CapG protein expression and localization in Sβtet29Cap35 (C35), Sβtet29Cap43 (C43) and the control Sβtet29 (TET29) cells was analyzed 24 h after treatment with 500 ng/ml doxycycline (+ dox) or PBS as control (- dox). Specific antibody reaction (anti-CapG) was visualized by a peroxidase labelled secondary antibody (DAB detection, brown colour). Nuclei were counterstained with hematoxylin (blue colour). Bars = 12.5 μm. B. Whole protein lysate (non frac.) or enriched nuclear (Nuc) and cytoplasmic (Cyto) fractions of Sβtet29Cap35, Sβtet29Cap43 and the control Sβtet29 clones 24 h after treatment with 500 ng/mL doxycycline (+) or PBS as control (-) were analysed using Western blotting for the detection of CapG, Lamin (marker for nuclear fraction) and GAPDH (marker for cytoplasmic fraction). The experiment was performed three times. A representative Western blot of Sβtet29Cap35 is shown.