Figure 4.

Gint4.T-STAT3 inhibits viability and migration of NSCLC cells co-cultured with CAFs

(A) Scheme of Calu-1 cells and CAFs co-cultures by Boyden chambers (pore size 0.4 μm) used for immunoblot and cell viability assays. (B and C) Calu-1 were co-cultured with CAFs in 5% FBS medium in the absence or in the presence of 400 nmol/L Gint4.T, Gint4.T-STAT3 or CtrlApt-STAT3 (see materials and methods for details) for 72 h. (B) Immunoblot for pSTAT3, STAT3 was performed. βActin or Vinculin antibodies were used as loading control. (C) Cells were recovered and cell viability was measured and expressed relative to cells grown in the absence of CAFs (−). (D) Scheme of Calu-1 cells and CAFs co-cultures using Boyden chambers (pore size 8 μm) to test cell migration. (E and F) Calu-1 cells were left untreated or treated with 400 nmol/L Gint4.T, Gint4.T-STAT3, or CtrlApt-STAT3 and following 24 h were seeded in the Boyden upper chamber and exposed to 1% FBS medium (−) or to CAFs grown in 1% FBS medium (lower chamber). Left panels, representative photographs of migrated cells are shown; right panels, migrated cells were quantified and expressed relative to (−) samples. In (C), (E), and (F), error bars depict SD values (n = 2). Statistics by t test are reported: ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001.