Figure 3. eCB enhancement reduces SM levels in cultured ASM‐KO neurons.

• A
  Mean ± SEM SM levels, expressed as percentage of vehicle values, in cultured neurons from ASM‐KO mice treated with vehicle or with the indicated concentrations of AEA (***P < 0.0001, n = 3 independent cultures, one‐way ANOVA, Bonferroni post hoc).
• B
  Mean ± SEM SM levels, expressed as percentage of vehicle values, in cultured neurons from ASM‐KO mice treated with vehicle, the inhibitor of NSM GW, AEA, or the combination of GW and AEA (***P < 0.0001, n = 3 independent cultures, one‐way ANOVA, Bonferroni post hoc).
• C
  Western blot against NSM and GAPDH (used as loading control) and graph showing mean ± SEM NSM protein levels in extracts from cultured neurons treated with vehicle or with 50 μM AEA (*P = 0.0230, n = 6 independent cultures, Student's t‐test).
• D
  Mean ± SEM SM levels, expressed as percentage of vehicle values, in cultured neurons from ASM‐KO mice treated with vehicle, or with JNJ, PF, or URB in the presence or absence of AEA (**P _JNJ = 0.0013, *P _PF = 0.0143, ***P _URB = 0.0002, **P _JNJ + AEA = 0.0069, **P _PF + AEA = 0.0077, ***P _URB + AEA < 0.0001, n = 3 independent cultures, one‐way ANOVA, Bonferroni post hoc).
• E
  Mean ± SEM SM levels, expressed as percentage of vehicle values, in cultured neurons from ASM‐KO mice treated with vehicle, with PF, or with SR141716 + PF (*P _PF = 0.0109, *P _SR + PF = 0.0343, n = 3 independent cultures, one‐way ANOVA, Bonferroni post hoc).
• F
  Immunofluorescences against the dendritic marker MAP2 of cultured neurons from ASM‐KO mice treated with vehicle, AEA, JNJ, PF, or URB. Scale bar, 50 μm.
• G
  Mean ± SEM number of apoptotic cells measured by TUNEL assays in cultured neurons from ASM‐KO mice treated with H₂O₂ (as positive control), vehicle, AEA, JNJ, PF, or URB (n = 3 independent cultures).

Source data are available online for this figure.