Fig. 8

EphrinB1-Fc treatment activates PI3K/Akt signaling in cultured Müller cells. a, Representative immunoblots showing the changes in PI3K p85 and p110 protein levels in IgG-Fc-treated cells (Ctr), and those treated with ephrinB1-Fc for different periods of time. b-c, Bar chart summarizing the average densitometric quantification of immunoreactive bands of PI3K p85 (b) and PI3K p110 (c) under different conditions as shown in A. n = 4 for all groups. All the data are normalized to Ctr. * P < 0.05, ** P < 0.01 and *** P < 0.001 vs. Ctr. d, Representative immunoblots showing that pre-incubation with RO25–6981 eliminated the ephrinB1-Fc-treatment induced changes in PI3K p85 and p110 protein levels in Müller cells. RO25–6981 (0.5 μM/5 μM) was added to the medium 30 min before a 6 h ephrinB1-Fc treatment. n = 5 for all groups. e-f, Bar chart summarizing the average densitometric quantification of immunoreactive bands of PI3K p85 (e) and p110 (f) under different conditions as shown in D. n = 5 for all groups. *P < 0.05, ** P < 0.01 and *** P < 0.001 vs. Ctr (IgG-Fc treated group). g, Representative immunoblots showing the changes in p-Akt and Akt protein levels in IgG-Fc-treated cells (Ctr), and those treated with ephrinB1-Fc for different periods of time. h, Bar chart summarizing the average p-Akt/Akt ratios in Müller cells treated with ephrinB1-Fc for different periods of time. n = 5 for all groups. * P < 0.05, vs. Ctr. i, Representative immunoblots showing that ephrinB1-Fc treatment induced changes in p-Akt and Akt protein levels were blocked by adding PP2 (10 μM) to the medium 30 min before a 3 h ephrinB1-Fc treatment. j, Bar chart summarizing the average p-Akt/Akt ratios of Müller cells under different conditions as shown in I. n = 5 for all groups. ** P < 0.01 vs. Ctr (IgG-Fc treated group)