Molecular Mechanisms of Tau Hyperphosphorylation

Unless indicated, all chemicals were purchased from Sigma/Merck Millipore. Antibodies used for immunoblotting included: mouse anti‐demethylated PP2Ac (clone 1D6, Merck Millipore); mouse anti‐PP2Ac_α/β (clone 46, BD Transduction); rabbit anti‐phospho‐p44/42 mitogen activated protein kinase (MAPK) (#9101, Cell Signaling Technology); mouse anti‐p44/42 MAPK (ERK1/2) (L34F12, Cell Signaling Technology); rabbit anti‐phospho‐GSK3β (Ser9) (D85E12, Cell Signaling Technology); mouse anti‐GSK3β (clone 3D10, Cell Signaling Technology); rabbit anti‐Tau (T‐1308‐1, rPeptide); mouse anti‐pS202‐Tau (CP13) [41 (link)] and anti‐pSer396/Ser404 (PHF‐1) tau (kindly donated by Peter Davies); mouse anti‐pS422‐tau (44‐764G, ThermoFisher); mouse anti‐human Tau‐13 (ab19030, Abcam); rabbit anti‐Fyn (clone EPR5500, Merck Millipore); mouse anti‐Fyn clone 25 (BD Transduction Laboratories); rabbit anti‐phospho‐SFK (Y416) (clone 100F9, Cell Signaling Technology); mouse anti‐actin (clone C4, Merck Millipore); and mouse anti‐glyceraldehyde 3‐phosphate dehydrogenase (clone 6C5, Merck Millipore). Mouse anti‐Fyn (clone 15, Santa Cruz), rabbit anti‐pS202‐Tau (ab108387, Abcam) and PHF‐1 antibodies were used for immunostaining.

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van Hummel A., Taleski G., Sontag J., Feiten A.F., Ke Y.D., Ittner L.M, & Sontag E. (2023). Methyl donor supplementation reduces phospho‐Tau, Fyn and demethylated protein phosphatase 2A levels and mitigates learning and motor deficits in a mouse model of tauopathy. Neuropathology and Applied Neurobiology, 49(4), e12931.

Publication 2023

mouse anti‐p44/42 MAPK (ERK1/2) (L34F12, rabbit anti‐phospho‐GSK3β (Ser9) (D85E12, mouse anti‐GSK3β (clone 3D10, rabbit anti‐Fyn (clone EPR5500, rabbit anti‐phospho‐SFK (Y416) (clone 100F9, mouse anti‐actin (clone C4, ab108387

Corresponding Organization : University of Newcastle Australia

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Demethylated PP2Ac
PP2Ac (α/β)
Phospho-p44/42 MAPK
P44/42 MAPK (ERK1/2)
Phospho-GSK3β (Ser9)
GSK3β
Phospho-Tau (pS202, pSer396/Ser404, pS422)
Phospho-SFK (Y416)
Actin
GAPDH

control variables

None explicitly mentioned

controls

Positive control: None explicitly mentioned
Negative control: None explicitly mentioned

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