Fig. 5

Impact of TSPAN3 expression on the intracellular recycling of β1 integrin. A, B β1 integrin (ITGB1) mRNA levels were detected through RT-PCR in A549 and H460 cells with tetraspanin 3 (TSPAN3/T3) knockdown and overexpression (n = 3 independent experiments). C Relative mRNA expression of TSPAN3 and ITGB1 in 16 NSCLC samples as analyzed by RT-PCR. The 2^−ΔΔCt value of each sample was calculated with the no. 6 sample as the reference. (P > 0.05). D Model depicting the cellular response associated with nocodazole (NZ) washout [2]. E β1 integrin expression on the surface of A549 cells at the indicated time points after NZ washout was assessed by flow cytometry. The expression values were normalized to those at 0 min (n = 3 independent experiments). F Surface expression of β1 integrin at 100 min after NZ washout relative to that at 60 min after NZ washout in A549 cells transfected with either si-TSPAN3 or si-Ctrl (n = 3). G The levels of p-FAK at different time points after NZ washout. The levels of p-FAK at 100 min after NZ washout relative to that at 60 min were calculated after normalizing to Lamin B levels (right, n = 3 independent experiments). H Immunofluorescence depicting the localization of TSPAN3 (TRITC) and β1 integrin (FITC) in A549 cells at different time points after NZ washout. Magnification ×400. *P < 0.05; **P < 0.01; ***P < 0.001