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. 2011 Mar 9;301(1):C99–C105. doi: 10.1152/ajpcell.00151.2010

Fig. 5.

Fig. 5.

Transfected N-cadherin fails to form cysts in 3D culture while cadherin 8 does. HK-2 cells were transfected with Flag-tagged N-cadherin or cadherin 8 for 1 day and then put into a 3D collagen matrix. A: immunoblot of Flag-tagged N-cadherin and cadherin 8. Lysates were made from HK-2 cells 24 and 48 h posttransfection. Blot was developed using anti-Flag tag antibody. Lanes 1 and 4, mock-transfected cells; lanes 2 and 4: lysates made from cells 24 h posttransfection; lanes 3 and 6: lysates made from cells 48 h posttransfection. At both time points, 24 and 48 h after transfection, significant amounts of Flag-tagged cadherin transgene are expressed. A, bottom: blot was stripped and probed with anti-actin. B: phase-contrast image of cells transfected with flag tagged N-cadherin grown in 3D culture 48 h postplating. Extensive cell chords and tubules were observed. Bar, 100 μm. C: volume rendering of 3D culture labeled with Hoescht 33342 (gray) and phalloidin (Texas red, red). Cells were transfected with N-cadherin. D: phase-contrast image of cells transfected with Flag-tagged cadherin 8. In addition to chords and tubules, cysts were evident in the 3D culture (arrows). Bar, 100 μm. E: volume rendering of 3D culture labeled with Hoescht 33342 (gray) and phalloidin (Texas red, red). Cells were transfected with cadherin 8. Bar, 100 μm.