Co-Immunoprecipitation of MeCP2 and CREB1

Co‐IP assay was performed in accordance with our previous study(Huang et al., 2019 (link)). In brief, after the supernatants of brain samples were collected, the lysates were incubated with MeCP2 antibody (Santa Cruz Biotechnology, Texas, USA) and protein A/G agarose gel overnight, followed by SDS‐PAGE resolution and Western blot assay. CREB1 antibody (Cell Signaling Technology, Inc., Danvers, USA) was used to measure the interaction of the two proteins.

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Huang J., Zhang F., Su M., Li J., Yi W., Hou L., Yang S., Liu J., Zhang H., Ma T, & Wu D. (2021). MeCP2 prevents age‐associated cognitive decline via restoring synaptic plasticity in a senescence‐accelerated mouse model. Aging Cell, 20(9), e13451.

Publication 2021

MeCP2 antibody CREB1 antibody

Agarose Antibody Assay Brain Mecp2 Protein g Proteins Sds page Western blot assay

Corresponding Organization : Nanjing Medical University

Other organizations : Guangxi University of Chinese Medicine

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Variable analysis

independent variables

Incubation of brain sample lysates with MeCP2 antibody (Santa Cruz Biotechnology, Texas, USA) and protein A/G agarose gel overnight

dependent variables

Interaction of MeCP2 and CREB1 proteins, measured by Western blot assay using CREB1 antibody (Cell Signaling Technology, Inc., Danvers, USA)

control variables

Previous study (Huang et al., 2019) protocol, which the current study is performed in accordance with

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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