Fig. 3

The combination of PTX and CK can affect JAK–STAT pathway. The Lewis cells were cocultured with SPC and treated with 60 μM CK, 10 nM PTX, or both for 48 h. All experiments were carried out in the co-culture system, including Lewis cells, SPC or the whole system. A The concentration of IL-2 and IL-6 in co-culture system was determined by ELISA. B The expression of IL-6R in Lewis cells and SPC was detected by Westren blot, β-actin was used as loading control. C, D The expression of JAK1/2, p-JAK1/2, STAT3/5 and p-STAT3/5 in SPC was detected by Western blot, β-actin was used as loading control. E, F The levels of p-STAT3 and p-STAT5 in both cytoplasm/membrane and nuclei fraction of SPC were determined by Western blot, β-actin and LaminA/C were used as loading control. G The immunofluorescence analysis performed with anti-p-STAT3 (red) and DAPI, Scale bar: 10 μm. H The immunofluorescence analysis performed with anti-p-STAT5 (red) and DAPI, Scale bar: 10 μm. Data are mean ± SD for ≥ 3 independent experiments; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, vs Ctrl