GHSR and DRD1 Protein Interaction Detection

Protein interactions between GHSR/DRD1 in mouse brain slices were detected using Duolink PLA detection kits (Sigma-Aldrich, DUO92008) following manufacturer’s instructions. The following primary antibodies were used: GHS-R1a (Santa Cruz Biotechnology, sc-10359, 1:100) and anti-DRD1 (Abcam, ab81296, 1:200). The specificity of antibodies to GHSR and DRD1 was validated as previously described (17 (link)). The following Duolink in Situ PLA Probes were used: anti–rabbit PLUS (Sigma-Aldrich, DUO92002) and anti–goat MINUS (Sigma-Aldrich, DUO92006). Images were collected on a Nikon Ti2 confocal microscope. The mean intensity of PLA⁺ signals was analyzed using Nikon-Elements Advanced Research software.

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Tian J., Guo L., Wang T., Jia K., Swerdlow R.H., Zigman J.M, & Du H. (2023). Liver-expressed antimicrobial peptide 2 elevation contributes to age-associated cognitive decline. JCI Insight, 8(10), e166175.

Publication 2023

GHS-R1a anti–rabbit PLUS anti–goat MINUS Nikon Ti2 confocal microscope

Antibodies Antibodies specificity Brain Confocal microscope Drd1 Ghs r1a Ghsr protein Goat Mouse Rabbit

Corresponding Organization : University of Kansas Medical Center

Other organizations : The University of Texas Southwestern Medical Center

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Variable analysis

independent variables

Use of Duolink PLA detection kits (Sigma-Aldrich, DUO92008) following manufacturer's instructions

dependent variables

Mean intensity of PLA+ signals

control variables

Primary antibodies used: GHS-R1a (Santa Cruz Biotechnology, sc-10359, 1:100) and anti-DRD1 (Abcam, ab81296, 1:200)
Duolink in Situ PLA Probes used: anti–rabbit PLUS (Sigma-Aldrich, DUO92002) and anti–goat MINUS (Sigma-Aldrich, DUO92006)
Images collected on a Nikon Ti2 confocal microscope
The specificity of antibodies to GHSR and DRD1 was validated as previously described (17 (link))

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