Fig. 7

EphrinB1-Fc treatment induces an increase in p-NR2B protein levels and increases TNF-α mRNA and protein levels in cultured Müller cells. a, Representative immunoblots showing the protein levels of NR1 in IgG-Fc-treated cells (Ctr), and those treated with ephrinB1-Fc for different periods of time. b, Bar chart summarizing the average densitometric quantification of immunoreactive bands of NR1. n = 4 for all groups.c, Representative immunoblots showing the protein levels of phosphorylated NR2B (p-NR2B) and NR2B in Müller cells treated with ephrinB1-Fc for different periods of time. d, Bar chart summarizing the average p-NR2B/NR2B ratios in Müller cells treated with ephrinB1-Fc for different periods of time. n = 6~ 7. * P < 0.05 and ** P < 0.01 vs. Ctr. e, Representative immunoblots showing that pre-incubation with PP2 eliminated the ephrinB1-Fc-treatment induced changes in p-NR2B protein levels in Müller cells. PP2 (10 μM) was added to the medium 30 min before a 3 h ephrinB1-Fc-treatment. f, Bar charts summarizing the average p-NR2B/NR2B ratios obtained in the presence of ephrinB1-Fc, with or without 30 min pre-incubation of 10 μM PP2. All the data are normalized to the ratio obtained in IgG-Fc treated group (Ctr). n = 4 for all groups. *** P < 0.001 vs. IgG-Fc treated group (Ctr). g, Summary data showing that relative TNF-α mRNA levels in Müller cells, represented as fold changes, were significantly elevated by ephrinB1-Fc treatment, but the elevation was abolished by addition of RO25–6981 (5 μM) 30 min before the ephrinB1-Fc treatment. n = 5 for all groups. ** P < 0.01 vs. Ctr (IgG-Fc treated group). h, Summary data showing that the ephrinB1-Fc treatment induced increase in TNF-α protein levels of cultured Müller cells was blocked by adding RO25–6981 (5 μM) to the medium 30 min before the ephrinB1-Fc treatment. n = 5 for all groups. *** P < 0.001 vs. Ctr (IgG-Fc treated group)