Figure 2-.
(F) Immunostaining of Hochest and GFP in cultured hippocampal neurons after GFP–PIAS1WT transfection. Cells that show both green fluorescence (GFP) and blue fluorescence (Hochest) are cells successfully transfected with the plasmid. Scale bar equals 25 μm. (G) Representative gel pattern and statistics for STAT1 sumoylation, STAT1 DNA binding and anti-Flag (verification of transfection) after Flag–PIAS1WT transfection and probe trial test. (H) Representative gel pattern and statistics for STAT1 phosphorylation at Tyr-701 after Flag–PIAS1WT transfection and probe trial test. N=8–10 each group. (I) DNA gel electrophoresis showing the GFP fragment (129 bp) from Q–PCR analysis after GFP–PIAS1WT transfection to rat CA1 area. Control animals received PEI (the transfection reagent) injection only. The primers used for Q–PCR analysis of GFP–PIAS1 mRNA level were designed within the sequence of GFP (upper panel). Quantitative analysis and statistics showing the effect of GFP–PIAS1WT transfection on GFP–PIAS1 mRNA expression (lower panel). N=6 each group. Data are mean±s.e.m. *P<0.05, **P<0.01 and ***P<0.001 from Student's t-test.