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. 2018 Oct 29;123(2):e239–e248. doi: 10.1016/j.bja.2018.08.025

Fig 3.

Fig 3

Phosphorylation of GluN2B is required for modulation of neuropathic pain related to receptor tyrosine kinase-like orphan receptor ROR2. (a) Representative western blots showing the effect of ROR2 small interfering RNA (siRNA) on serine phosphorylation of N-methyl-D-aspartic acid (NMDA) receptors induced by chronic constriction injury (CCI). P-Ser, serine phosphorylation. (b) Quantitative data of serine phosphorylation of GluN1 subunit. *P<0.05, n=6. Scr, scramble. (c) Quantitative data of serine phosphorylation of GluN2A subunit. ***P<0.001, n=6. (d) Quantitative data of serine phosphorylation of GluN2B subunit. *P<0.05, **P<0.01, n=6. (e) Representative western blots showing the effect of ROR2 siRNA (10 μg in 0.5 μl) on tyrosine phosphorylation of NMDA receptors induced by CCI. (f) Quantitative data of tyrosine phosphorylation of GluN1 subunit. *P<0.05, n=6. (g) Quantitative data of tyrosine phosphorylation of GluN2A subunit. *P<0.05, n=6. (h) Quantitative data of tyrosine phosphorylation of GluN2B subunit. *P<0.05, ***P<0.001, n=6. Phosphorylation of NMDA receptor was assessed on Day 11, the same as in Figure 2d. (i) Representative western blots and quantitative data for phosphorylation of GluN2B subunit at Ser 1303, Ser 1323, Tyr1336, and Tyr1472 sites after ROR2 siRNA (10 μg in 0.5 μl) or scrambled siRNA injection. *P<0.05, **P<0.01, n=6. (j) Representative western blots and quantitative data for phosphorylation of GluN2B subunit at Ser1303 and Tyr1472 sites after Wnt5a (0.5 μg in 0.5 μl) or saline injection. *P<0.05, **P<0.01, n=6. (k) Effect of pretreatment with MK801(2.5 μg in 0.5 μl) and ifenprodil (0.5 μg in 0.5 μl) on thermal hyperalgesia and mechanical allodynia induced by Wnt5a. *P<0.05, **P<0.01, Saline-Wnt5a vs MK801-Wnt5a; #P<0.05, ##P<0.01, Saline-Wnt5a vs Ifenprodil-Wnt5a; n=8. Ifen, ifenprodil. (l) Effect of posttreatment with MK801 (2.5 μg in 0.5 μl) and ifenprodil (0.5 μg in 0.5 μl) on thermal hyperalgesia and mechanical allodynia induced by Wnt5a. *P<0.05, **P<0.01, ***P<0.001, Wnt5a-Saline vs Wnt5a-MK801; #P<0.05, ##P<0.01, ###P<0.01, Wnt5a-Saline vs Wnt5a-Ifenprodil; n=8.