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. 2001 Sep;12(9):2730–2741. doi: 10.1091/mbc.12.9.2730

Figure 7.

Figure 7

Stable transfection of 3T3 fibroblasts with different actin isoforms. 3T3 fibroblasts (A) were stably transfected with VSV-G–tagged α-SMA (B), γ-cytoplasmic (C) and α-cardiac actin (D) cDNA, respectively. Double-immunostaining of wild-type 3T3 fibroblasts demonstrated low expression of α-SMA (A, red) in F-actin–containing stress fibers (A, green) Transfected α-SMA and α-cardiac actin were predominantly incorporated into stress fibers and γ-cytoplasmic actin into membrane ruffles and lamellipodia, as shown by double-immunostaining of fibroblasts for actin isoforms (B–D, green) and VSV-G-tag (B–D, red). Bar, 50 μm. Western blotting (E) demonstrated low α-SMA expression in wild-type 3T3 fibroblasts (3T3wt, co) compared with TGFβ1-treated 3T3 fibroblasts (3T3wt, TGF) and LFs. Note that enhanced expression of α-SMA correlates with enhanced VSV-G-tag expression in α-SMA transfected clones (s1–s3), but not in clones expressing transfected α-cardiac (α-Car: c1, c2) and γ-cytoplasmic actin (γ-Ccyt: y1, y2). Vimentin was used to demonstrate equal loads of total protein.