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. 2020 May 19;11:2501. doi: 10.1038/s41467-020-16198-5

Fig. 8. nNOS-expressing neurons in the vmPFC mediate chronic pain-induced anxiety through diffusing NO.

Fig. 8

a, b Dot plot (a, upper) and western bolts (a, lower) showing nNOS protein levels (n = 4), and dot plot (b) showing nNOS enzyme activity in the vmPFC at day 3 after CFA or NS injection (n = 6). c Dot plot showing nNOS enzyme activity of the vmPFC 30 min after CNO (0.5 mM, 1 μl) or NS delivery at day 21 after AAV-hSyn-DIO-hM3Dq-eGFP microinjection (0.4 μl) in the vmPFC of nNOS-Cre mice (n = 5). d–f Distance in center (d, left) and total distance (d, right) in an OF test, time in open arms (e, left) and total entries in arms (e, right) of an EPM test, and withdrawal threshold of hind paw in an von Frey test (f) 30 min after L-VNIO (1.5 mM, 1 μl) or NS (1 μl) microinjection into vmPFC at day 3 after CFA or NS injection. n = 15. g–i Distance in center (g, left) and total distance (g, right) with an OF test, time in open arms (h, left) and total entries in arms (h, right) of an EPM test, and withdrawal threshold of the hind paw in an von Frey test (i) 30 min after L-VNIO (1.5 mM, 1 μl) or NS (1 μl) microinjection in vmPFC at day 7 after SNL or sham operation. n = 13, 14, and 12, respectively. j, k Distance in center (j, left) and total distance (j, right) with an OF test, and time in open arms (k, left) and total entries in arms (k, right) of an EPM test 30 min after L-VNIO (1.5 mM, 1 μl) or NS microinjection in vmPFC. AAV-hSyn-DIO-hM3Dq-eGFP (0.4 μl) was microinjected into the vmPFC, and 3 weeks later, CNO (0.5 mM, 1 μl) was microinjected into the vmPFC 30 min before L-VNIO or NS microinjection. n = 15. Data are the mean ± SEM; *p < 0.05, **p < 0.01, and ***p < 0.001 (unpaired two-tailed Student’s t-test for (a–c) and (j), (k); one-way ANOVA followed by Tukey’s post hoc test for (d–i)). Source data are provided as a Source data file. Exact p values and additional statistical information can be found in Source data.