The knocking down of GOLPH3 affects the reassembly of focal adhesion in T98G cells.

(A-F) The indicated cells grown in glass coverslips were treated with 10 μM nocodazole for 4 h at 37°C (A and B), or subsequently subjected to nocodazole washout for either 20 min (C and D) or 2-h (E and F) at 37°C. Cells were fixed, permeabilized, and double-labeled with mouse monoclonal antibody to vinculin, and TRITC-phalloidin, followed by Alexa-488-conjugated donkey anti-mouse IgG. Stained cells were examined by fluorescence microscopy. Bar, 10 μm. (G) Quantification of focal adhesions of the indicated cells subjected to the indicated treatments from images as shown in A-F, performed as indicated in Materials and Methods. Bar represents the mean ± standard deviation (n = 30 cells). * P P ns, not statistically significant.