Fig. 7.

Trans- and classic signalling-induced growth of Ba/F3-gp130-IL-6Rα cells do not differ. a Ba/F3-gp130-IL-6Rα cells were stimulated with IL-6 (blue) or Hy-IL-6 (red) as indicated. After 48 h of incubation cell growth was measured using Cell Titer Blue reagent. Diamonds correspond to the mean and bars to the standard deviation for n = 4 experiments. b Ba/F3-gp130-IL-6Rα cells were stimulated with IL-6 (blue) and Hy-IL-6 (red) (0.17 nM). STAT3 phosphorylation and STAT3 expression were evaluated by Western blotting. STAT3 expression served as loading control. Diamonds correspond to the mean and bars to the standard deviation for n = 4 independent experiments. Data normalization was performed as described in Additional file 1: Text S3. c Ba/F3-gp130-IL-6Rα cells were stimulated with the indicated amounts of IL-6 (blue bars) or Hy-IL-6 (red bars), respectively. STAT3 phosphorylation was evaluated by intracellular flow cytometry using specific fluorescent antibodies against STAT3 (p)Y705. For independent experiments mean fluorescence of 10⁴ cells per cytokine concentration was calculated. Data are given as mean ± STD from n = 3 experiments. d The expression of gp130 and IL-6Rα at the surface of Ba/F3-gp130-IL-6Rα cells was analysed by flow cytometry using QIFIKIT. Mean ± STD from n = 4 independent experiments is shown