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. 2008 Jun 25;105(26):8950–8955. doi: 10.1073/pnas.0711264105

Fig. 2.

Fig. 2.

A study by transient expression of laminin fragments in Sertoli cells to assess their effects on the BTB integrity and the steady-state levels of different target proteins. (A) RT-PCR and immunoblot analysis showing the expression of domain IV of laminin γ3 (Left), domain I of laminin γ3 (Center), and domain I of laminin β3 (Right) in Sertoli cells after transfection with laminin γ3 DIV/pCIneo, laminin γ3 DI/pCIneo, and laminin β3 DI/pCIneo expression vectors for 2 and 3 days, respectively, vs. cells transfected with pClneo alone (without laminin cDNA constructs) or normal cells (Ctrl). S16, GAPDH, and actin served as loading controls. (B) Changes in Sertoli cell–TJ barrier by quantifying TER across the cell epithelium in bicameral units (n = 3) where cells were transfected with laminin γ3 DIV/pCIneo vs. pCIneo alone on day 2.5 (arrowhead). (C) The protein levels of β1-integrin, occludin, and ERK/pERK in cells following overexpression with different constructs for 3 days vs. controls. (D–E) Bar graphs summarize results of C. Each bar is a mean ± SD (n = 3) and normalized against actin, wherein cells transfected with pCIneo vector alone were arbitrarily set at 1, against which one-way ANOVA was performed. ns, not significantly different; ∗, P < 0.05; ∗∗, P < 0.01.