Fig. 3
Knockdown of YTHDF1 in the vlPAG by CMV-Cre-EGFP in YTHDF1fl/fl mice decreased morphine analgesic tolerance and MIH. a, b Immunostaining verifying YTHDF1 downregulation in the vlPAG of CMV-Cre-EGFP in YTHDF1fl/fl mice. Scale bars, 50 μm. c Western blotting analysis verifying YTHDF1 downregulation in the vlPAG of CMV-Cre-EGFP in YTHDF1fl/fl mice (***P < 0.001, n = 6, Student’s t test, two-tailed). d, e Nociceptive behavior (BL time): hot plate, F1,18 = 4.87, P = 0.041; tail immersion, F1,18 = 20.69, P < 0.001. Antinociception: hot plate, F1,18 = 15.576, P = 0.001; tail immersion, F1,18 = 22.084, P < 0.001. f–g Morphine antinociception (%MPE). f Hot plate, left, F1,18 = 21.037, P < 0.0001; ***P < 0.001 vs. Day 1 + 30 min of the CMV-EGFP group; ###P < 0.001, vs. Day 1 + 30 min of the CMV-Cre-EGFP group; †††P < 0.001, vs. Day 7 + 30 min of the CMV-EGFP group; right, the percent change for each subject, ***P < 0.001 vs. the CMV-EGFP group; g Tail immersion, left, F1,18 = 11.197, P = 0.004; ***P < 0.001 vs. Day 1 + 30 min of the CMV-EGFP group; ###P < 0.001, vs. Day 1 + 30 min of CMV-Cre-EGFP group; †††P < 0.001, vs. Day 7 + 30 min of CMV-EGFP group; right, the percent change for each subject, ***P < 0.001 vs. the CMV-EGFP group; h, i changes in withdrawal latency before and after morphine administration. h Hot plate: left, F1,18 = 33.564, P < 0.001; ***P < 0.001 vs. Day 1 BL of the CMV-EGFP group; N.S. P = 0.0599 vs. Day 1 BL of the CMV-Cre-EGFP group; ###P < 0.001, vs. Day 7 BL of the CMV-EGFP group; right, the percent change for each subject, **P = 0.00962 vs. the CMV-EGFP group; i tail immersion: left, F1,18 = 67.778, P < 0.001; ***P < 0.001 vs. Day 1 BL of the CMV-EGFP group; N.S. P = 0.1045 vs. Day 1 BL of the CMV-Cre-EGFP group; ###P < 0.001, vs. Day 7 BL of the CMV-EGFP group; right, the percent change for each subject, ***P < 0.001 vs. the CMV-EGFP 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)