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The Regulatory Factor ZFHX3 Modifies Circadian Function in SCN via an AT Motif-Driven Axis.

Parsons MJ, Brancaccio M, Sethi S, Maywood ES, Satija R, Edwards JK, Jagannath A, Couch Y, Finelli MJ, Smyllie NJ, Esapa C, Butler R, Barnard AR, Chesham JE, Saito S, Joynson G, Wells S, Foster RG, Oliver PL, Simon MM, Mallon AM, Hastings MH, Nolan PM - Cell (2015)

Bottom Line: Using RNA sequencing, we found minimal effects on core clock genes in Zfhx3(Sci/+) SCN, whereas the expression of neuropeptides critical for SCN intercellular signaling was significantly disturbed.Lentiviral transduction of SCN slices showed that the ZFHX3-mediated activation of AT motifs is circadian, with decreased amplitude and robustness of these oscillations in Zfhx3(Sci/+) SCN slices.In conclusion, by cloning Zfhx3(Sci), we have uncovered a circadian transcriptional axis that determines the period and robustness of behavioral and SCN molecular rhythms.

View Article: PubMed Central - PubMed

Affiliation: MRC Harwell, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, UK.

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Validation of the ZFHX3 Antibody, Related to Experimental ProceduresExpression and characterization of ZFHX3 in mouse tissues.(A) Western blot analysis showing ZFHX3 immunoreactive product in mouse SCN, hypothalamus, and embryonic brain lysates.(B) Immunoprecipitation of ZFHX3 using anti-ZFHX3 antibody and validation by mass spectrometry.
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figs7: Validation of the ZFHX3 Antibody, Related to Experimental ProceduresExpression and characterization of ZFHX3 in mouse tissues.(A) Western blot analysis showing ZFHX3 immunoreactive product in mouse SCN, hypothalamus, and embryonic brain lysates.(B) Immunoprecipitation of ZFHX3 using anti-ZFHX3 antibody and validation by mass spectrometry.

Mentions: Confocal microscopy was used to image immunofluorescence in free-floating brain sections (40 μm) using a range of antibodies and quantified using ImageJ software (Figure S7; Supplemental Experimental Procedures).


The Regulatory Factor ZFHX3 Modifies Circadian Function in SCN via an AT Motif-Driven Axis.

Parsons MJ, Brancaccio M, Sethi S, Maywood ES, Satija R, Edwards JK, Jagannath A, Couch Y, Finelli MJ, Smyllie NJ, Esapa C, Butler R, Barnard AR, Chesham JE, Saito S, Joynson G, Wells S, Foster RG, Oliver PL, Simon MM, Mallon AM, Hastings MH, Nolan PM - Cell (2015)

Validation of the ZFHX3 Antibody, Related to Experimental ProceduresExpression and characterization of ZFHX3 in mouse tissues.(A) Western blot analysis showing ZFHX3 immunoreactive product in mouse SCN, hypothalamus, and embryonic brain lysates.(B) Immunoprecipitation of ZFHX3 using anti-ZFHX3 antibody and validation by mass spectrometry.
© Copyright Policy - CC BY
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4537516&req=5

figs7: Validation of the ZFHX3 Antibody, Related to Experimental ProceduresExpression and characterization of ZFHX3 in mouse tissues.(A) Western blot analysis showing ZFHX3 immunoreactive product in mouse SCN, hypothalamus, and embryonic brain lysates.(B) Immunoprecipitation of ZFHX3 using anti-ZFHX3 antibody and validation by mass spectrometry.
Mentions: Confocal microscopy was used to image immunofluorescence in free-floating brain sections (40 μm) using a range of antibodies and quantified using ImageJ software (Figure S7; Supplemental Experimental Procedures).

Bottom Line: Using RNA sequencing, we found minimal effects on core clock genes in Zfhx3(Sci/+) SCN, whereas the expression of neuropeptides critical for SCN intercellular signaling was significantly disturbed.Lentiviral transduction of SCN slices showed that the ZFHX3-mediated activation of AT motifs is circadian, with decreased amplitude and robustness of these oscillations in Zfhx3(Sci/+) SCN slices.In conclusion, by cloning Zfhx3(Sci), we have uncovered a circadian transcriptional axis that determines the period and robustness of behavioral and SCN molecular rhythms.

View Article: PubMed Central - PubMed

Affiliation: MRC Harwell, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, UK.

Show MeSH