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Fig. 5 | Acta Neuropathologica Communications

Fig. 5

From: EphrinB/EphB forward signaling in Müller cells causes apoptosis of retinal ganglion cells by increasing tumor necrosis factor alpha production in rat experimental glaucomatous model

Fig. 5

Activation of ephrinB/EphB forward signaling increases TNF-α production in cultured Müller cells and retinal tissues. The cells were treated by IgG-Fc (500 ng/ml) (Ctr) or ephrinB1-Fc (500 ng/ml) for different periods of time (1–24 h). a-c, Cumulative data summarizing the changes in mRNA levels of BDNF (a), GDNF (b) and NGF (c) in Müller cell extracts obtained in Ctr and those with ephrinB1-Fc-treatment for different periods of time. n = 6 for all groups. d-f, Cumulative data summarizing the changes in mRNA levels of IL-1β (d), IL-6 (e) and TNF-α (f) in Müller cell extracts obtained in Ctr and those with ephrinB1-Fc-treatment for different periods of time. The relative abundance of mRNA was quantified using 2-ΔΔct calculation method and expressed as fold change. n = 6 for all groups.* P < 0.05 and *** P < 0.001 vs. Ctr. g, Bar chart showing the average extracellular TNF-α concentrations in IgG-Fc-treated group (Ctr) and groups of ephrinB1-Fc-treatment for different periods of time. n = 6 for all groups. * P < 0.05 and *** P < 0.001 vs. Ctr. h, Summary data showing that the ephrinB1-Fc-treatment induced increase in TNF-α mRNA levels in cultured Müller cells was blocked by pre-incubation with PP2. PP2 (10 μM/50 μM) was added to the medium 30 min before a 6 h ephrinB1-Fc treatment. n = 6 for all groups. *** P < 0.001 vs. IgG-Fc treated group (Ctr). i, Bar chart showing that pre-incubation with PP2 inhibited the ephrinB1-Fc-treatment induced increase in TNF-α protein levels in cultured Müller cells. PP2 (10 μM) was added to the medium 30 min before a 24 h ephrinB1-Fc treatment. n = 6 for all groups. *** P < 0.001 vs. IgG-Fc treated group (Ctr). j, Cumulative data summarizing the changes in TNF-α mRNA levels in normal saline-injected retinas (Ctr) and retinas with ephrinB1-Fc-injection at different post-injection times. EphrinB1-Fc (0.5 μg/μl, 2 μl) was intravitreally injected and retinas were collected at different post-injection times (3 d and 7 d). n = 6 for all groups. k, Bar chart showing the changes in TNF-α protein levels in Ctr retinas and ephrinB1-Fc injected retinas at different post-injection times (3 d and 7 d). n = 4 for all groups. * P < 0.05 and *** P < 0.001 vs. Ctr

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