Expression of profilin 1 mutants in PC-3 cells.

<p>(A) Scheme of the DNA construct of profilin 1 mutants. (B) Restriction analysis of the DNA construct ligated into the pcDNA3 cloning vector using EcoRI and BamHI. Line 1: GeneRuler 1 kb DNA Ladder (Promega), line 2: restriction of Pfn1-Tyr139 mutant, line 3: restriction of Pfn1-Q138P mutant. (C) Western blot analysis of transfected PC-3 cell lysates expressing different profilin 1 mutants. Cell lysates were prepared 48 h post transfection. Line 1: size marker (SeeBlue® Pre-stained Protein Standard, Life Technologies), line 2: transfected empty pcDNA3 vector, line 3: transfected Pfn1-Tyr139/pcDNA3, line 4: transfected Pfn1-Q138P/pcDNA3. Mutants were detected with anti-FLAG antibodies. Detection of profilin 1 (native and mutated) and β-actin on the same membrane is also shown. Due to the chemiluminescent detection, size marker is added as a separate strip.