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Fig. 2 | Journal of Neuroinflammation

Fig. 2

From: The YTHDF1–TRAF6 pathway regulates the neuroinflammatory response and contributes to morphine tolerance and hyperalgesia in the periaqueductal gray

Fig. 2

Knocking down YTHDF1 expression in vlPAG by YTHDF1-shRNA decreased analgesic tolerance and MIH. a, b Immunostaining verifying YTHDF1 downregulation in the vlPAG of CMV-YTHDF1-shRNA-NC or CMV-YTHDF1-shRNA in WT mice. Scale bars, 50 μm. c Western blotting analysis verifying YTHDF1 downregulation in the vlPAG of CMV-YTHDF1-shRNA in WT mice (P = 0.0005, n = 6, Student’s t test, two-tailed). d, e Nociceptive behavior (BL time): hot plate, F1,18 = 0.553, P = 0.467; tail immersion, F1,18 = 2.158, P = 0.159. Antinociception: hot plate, F1,18 = 17,546, P = 0.001; tail immersion, F1,18 = 14.946, P = 0.001. f, g Morphine antinociception (%MPE). f Hot plate, left, F1,18 = 11.74, P = 0.003. *** P < 0.001 vs. Day 1 + 30 min of the shRNA-NC group; ### P < 0.001, vs. Day 1 + 30 min of the YTHDF1-shRNA group; †† P = 0.00126, vs. Day 7 + 30 min of the shRNA-NC group; right, the percent change for each subject, ** P = 0.00151 vs. the shRNA-NC group; g Tail immersion, left, F1,18 = 18.776, P < 0.0001. *** P < 0.001 vs. Day 1 + 30 min of the shRNA-NC group; ### P < 0.001, vs. Day 1 + 30 min of the YTHDF1-shRNA group; †† P = 0.001, vs. Day 7 + 30 min of the shRNA-NC group; right, the percent change for each subject, *** P < 0.001 vs. the shRNA-NC group; h, i Changes in withdrawal latency before and after morphine administration. h Hot plate: left, F1,18 = 20.108, ***P < 0.001 vs. Day 1 BL of the shRNA-NC group; N.S. P = 0.1422 vs. Day 1 BL of the YTHDF1-shRNA group; ##P = 0.00246, vs. Day 7 BL of the shRNA-NC group; right, the percent change for each subject, ***P < 0.001 vs. the shRNA-NC group; i Tail immersion: left, F1,18 = 24.516, P < 0.001. ***P < 0.001 vs. Day 1 BL of the shRNA-NC group; N.S. P = 0.1095 vs. Day 1 BL of the YTHDF1-shRNA group; #P = 0.0197, vs. Day 7 BL of shRNA-NC group; right, the percent change for each subject, *** P < 0.001 vs. shRNA-NC group; (repeated measures two-way ANOVA, n = 10) (Data are shown as the mean ± SEM. %MPE = percent of maximal possible effect; N.S. = No significance; CMV-YTHDF1-shRNA-NC = AAV-CMV-Cre + AAV-CMV-DIO-EGFP-YTHDF1-shRNA-NC; CMV-YTHDF1-shRNA = AAV-CMV-Cre + AAV-CMV-DIO-EGFP-YTHDF1-shRNA)

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