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. 2005 Dec 5;171(5):799–810. doi: 10.1083/jcb.200506083

Figure 6.

Figure 6.

Endogenous nesprin-3 and plectin associate in TM-4 cells. (A) The mouse Sertoli cell line TM-4 was transiently transfected with either the pSUPER vector expressing the nesprin-3 siRNA (A–D) or the luciferase siRNA (E–H), along with an expression vector for mRFP at a 5:1 ratio. After 72 h, the cells were fixed and stained for endogenous plectin (A and E) and nesprin-3 using our pAbs (C and G). mRFP was visualized to determine which cells express the siRNAs (B and F). D and H are composite images of the first three images in each row. All images are maximum projections. Bar, 20 μM. (B) TM-4 cells were left untransfected (A–H) or transiently transfected with the pSUPER vector expressing the nesprin-3 siRNA, along with an expression vector for mRFP in a 5:1 ratio (I–L). After 72 h and just before fixation, the cells in images E–L were treated for 30 min with 0.2 μM latrunculin B. The cells were then fixed and stained for plectin (B, F, and J), nesprin-3 (C, G, and K), and F-actin (A and E). mRFP was visualized to determine which cells express the nesprin-3 siRNA (I). D, H, and L are composite images of the first three images in each row. Bar, 10 μM.