Fig 6 Fluorescent immunostaining of the signal corresponding

Fig. 6. Fluorescent immunostaining of the signal corresponding to staining with N-PAT 1 and an antibody against GAPDH in A 7 r 5 cells. Cells were transfected with a solution of specific si. RNAs directed against PAT 1 or a solution of si. RNAs directed against GAPDH using the Hiperfect transfection procedure. A: cells were fixed on permeable polycarbonate filters 72 h after the 1 st of 2 transfections 72 and 24 h preevaluation. The cell nucleus is visualized with propidium iodide. Scale bars are 20 μm, and all sections are acquired under similar laser intensity to be comparable for quantification. Immunocytochemical staining after transient transfection with Hiperfect was repeated with 2 different passages with similar results. B: cell proliferation as a function of time in untreated A 7 r 5 cells (normal), A 7 r 5 cells transfected with a noncoding si. RNA ( GFP-si. RNA) and si. RNA directed toward PAT 1 (PAT 1 -si. RNA) or GAPDH (GAPDH-si. RNA). Results are reported as means ± SE of 3 different experiments. Significant difference in growth rate could be observed between GFPsi. RNA and PAT 1 -si. RNA at 96 h after transfection using one-way ANOVA followed by Tukey's posttest. * P < 0. 05. C: Western blot analysis of PAT 1 and GAPDH protein levels in confluent and dividing A 7 r 5 cells and confluent Caco-2 cells. DOI: (10. 1152/ajpendo. 00322. 2013)