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. Author manuscript; available in PMC: 2023 Apr 17.
Published in final edited form as: Mol Psychiatry. 2022 Aug 26;27(12):5213–5226. doi: 10.1038/s41380-022-01750-0

Fig. 7. Tau is a coregulator for KDM6B to control VGLUT1 and VGLUT2 expression.

Fig. 7.

(A) Co-IP with anti-Flag antibody in HEK293T cells transfected with Flag-KDM6B-C and Myc-Tau. WCL, whole cell lysate. (B) Co-IP with anti-Myc antibody in HEK293T cells transfected with Myc-Tau and Flag-KDM6B-C. (C) qRT-PCR analysis of Tau, VGLUT1, and VGLUT2 in neurons infected with lentivirus encoding SC shRNA or Tau shRNA1 (sh1) or shRNA2 (sh2). n = 4 biological replicates. One-way ANOVA with Dunnett’s multiple comparison test. (D, E) Immunoblot analysis of Tau, H3K27me3, VGLUT1 and VGLUT2 proteins in neurons infected with lentivirus encoding SC shRNA or Tau sh1 or sh2 (D). The intensity of Tau, H3K27me3, VGLUT1, and VGLUT2 bands was quantified and normalized to β-actin (E). n = 3 biological replicates. One-way ANOVA with Dunnett’s multiple comparison test. (F-I) Immunoblot analysis of KDM6B-C, Tau, H3K27me3, VGLUT1 and VGLUT2 proteins in WT and KDM6B KO neurons expressing Tau shRNA and/or Flag-KDM6B-C (F). The intensity of VGLUT1 (G), VGLUT2 (H), and Tau (I) bands was quantified and normalized to β-actin. n = 3 biological replicates. One-way ANOVA with Tukey’s multiple comparison test. (J, K) ChIP-qPCR analysis of KDM6B enrichment at the promoters of Slc17a7 (J) and Slc17a6 (K) in WT and Tau KD neurons expressing Flag-KDM6B-C. n = 3 biological replicates. Two-way ANOVA with Tukey’s multiple comparison test. (L, M) ChIP-qPCR analysis of H3K27me3 enrichment at the promoters of Slc17a7 (L) and Slc17a6 (M) in WT and Tau KD neurons. n = 3 biological replicates. Two-way ANOVA with Tukey’s multiple comparison test. Data were presented as mean ± s.e.m. **p < 0.01. ***p < 0.001; ****p < 0.0001.