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Fig. 7 | Journal of Neuroinflammation

Fig. 7

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

Fig. 7

YTHDF1 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)

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