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. 2016 May 27;6:26597. doi: 10.1038/srep26597

Figure 4. Knockdown of Ash1L reduced neuronal activity-induced repression of nrxn1α.

Figure 4

(Α) Τhe design of doxycycline-inducible shRNAmir against Ash1L. The transcription of shRNAmir and TurboRFP are under the control of the tetracycline response element (TRE) promoter, which can be activated by the reverse tetracycline transactivator 3 (rtTA3) in the presence of Dox. (B,C) Reduction of mRNA Ash1L (n = 3 each group; Student’s t-test, ***P < 0.001) and protein levels (full-length blots in Fig. S4; n = 4 each group; Student’s t-test, *P < 0.05) 48 hours after induction in NG108-15 cells. (D–F) Optogenetic stimulation induced expression changes of egr1 (E) and nrxn1α (F) in differentiated NG108-15 cells. Real-time PCR was performed for quantification (n > 6 biological samples, one-way ANOVA with Dunnett’s post hoc test, *P < 0.05, ***P < 0.001). (G) The activity-dependent repression of nrxn1α was rescued by the Dox-induced knockdown of Ash1L knock-down cells (n > 5; two-way ANOVA with Bonferroni post hoc test, *P < 0.05, **P < 0.01, ***P < 0.001).