Epigenetic Regulation of Neural Circuits

The mice were euthanized, and the brain tissue was harvested as indicated by the conditions. Immunofluorescence staining, western blotting and chromatin immunoprecipitation were performed as previously described (Lee et al., 2015 (link); Shi et al., 2013 (link); Xu et al., 2007 (link)). The antibodies used were the following: mouse anti-ZBTB20 9A10 (1:1,000, home made), mouse anti-BMAL1 (1:1,000, home made), goat anti-PROKR2 (Santa Cruz Biotechnology, 1:400, Cat# sc-54317 RRID:AB_2253158), rabbit anti-AVP (Millipore,1:4000, Cat# AB1565 RRID:AB_11212336); rabbit anti-VIP (Immunostar, 1:2000, Cat# 20077 RRID:AB_572270); rabbit anti-P300 (Santa Cruz Biotechnology, 1:400, Cat# sc-584 RRID:AB_2293429); mouse anti-NeuN (Millipore, 1:100, Cat# MAB377 RRID:AB_2298772); goat anti-GAD65/67 (Santa Cruz Biotechnology, 1:400, Cat# sc-7513 RRID:AB_2107745); rabbit anti-acetyl-histone H3 (Lys9) (Millipore, 1:500, Cat# 07–352 RRID:AB_310544); and rabbit anti-dimethyl-histone H3 (Lys9) (Upstate Biotechnology, 1:500, Cat# 07–441 RRID:AB_310619). The NIH Image J (RRID:SCR_003070) software was used for the quantification of the band from Western blots. The primer sequences for CHIP are provided in Figure 5—source data 1.

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Qu Z., Zhang H., Huang M., Shi G., Liu Z., Xie P., Li H., Wang W., Xu G., Zhang Y., Yang L., Huang G., Takahashi J.S., Zhang W.J, & Xu Y. (2016). Loss of ZBTB20 impairs circadian output and leads to unimodal behavioral rhythms. eLife, 5, e17171.

Publication 2016

rabbit anti-VIP rabbit anti-P300 mouse anti-NeuN rabbit anti-acetyl-histone H3 (Lys9)

Antibodies Brain Chip Chromatin immunoprecipitation Gad65 Goat Histone h3 Immunofluorescence Mouse P300 Primer Rabbit Tissue Western blots Zbtb20

Corresponding Organization :

Other organizations : Nanjing University, Model Animal Research Center, Second Military Medical University, Soochow University, The University of Texas Southwestern Medical Center

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Variable analysis

independent variables

Euthanasia of mice
Harvesting of brain tissue

dependent variables

Immunofluorescence staining
Western blotting
Chromatin immunoprecipitation

control variables

Antibodies used for immunofluorescence staining and western blotting, including: mouse anti-ZBTB20 9A10, mouse anti-BMAL1, goat anti-PROKR2, rabbit anti-AVP, rabbit anti-VIP, rabbit anti-P300, mouse anti-NeuN, goat anti-GAD65/67, rabbit anti-acetyl-histone H3 (Lys9), and rabbit anti-dimethyl-histone H3 (Lys9)

controls

Positive controls: Not explicitly mentioned.
Negative controls: Not explicitly mentioned.

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