Fig. 7From: The YTHDF1–TRAF6 pathway regulates the neuroinflammatory response and contributes to morphine tolerance and hyperalgesia in the periaqueductal grayYTHDF1 regulates the expression of TRAF6 in the vlPAG during morphine analgesic tolerance and MIH. Immunostaining verifying that knockdown of YTHDF1 in the vlPAG by CMV-YTHDF1-shRNA in WT mice (a–b) or CMV-Cre-EGFP in YTHDF1fl/fl mice (d–e) can inhibit the expression of TRAF6. Scale bars, 50 μm. c, f Western blotting analysis verifying that knockdown of YTHDF1 in the vlPAG by CMV-YTHDF1-shRNA in WT mice (c, **P = 0.00738, n = 6)) or CMV-Cre-EGFP in YTHDF1fl/fl mice (f, ***P < 0.0001, n = 6) inhibited the expression of TRAF6. g Summary of Traf6 mRNA measured in the vlPAG by chronic morphine treatments expressing CMV-YTHDF1-shRNA-NC or CMV-YTHDF1-shRNA, CMV-EGFP or CMV-Cre-EGFP relative to GAPDH mRNA (left, N.S. P = 0.430, n = 4; right, N.S. P = 0.7133, n = 4). h MeRIP-qPCR analysis of the m6A levels of TRAF6 mRNA in vlPAG with chronic saline or morphine treatments. (**P = 0.00133 vs. the control group) (n = 6, Student’s t test, two-tailed. Data are shown as the mean ± SEM, 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)Back to article page