. Author manuscript; available in PMC: 2019 Mar 25.

Published in final edited form as: Cell Rep. 2019 Jan 29;26(5):1089–1097.e3. doi: 10.1016/j.celrep.2019.01.004

Table 1.

Both LTD Induction by Glutamate + Depolarization Pairing and Transfection with a Tamoxifen-Inducible MEF2 Construct Activated MEF2 Transcriptional Activity as Measured Using an MEF2 Reporter Plasmid

Manipulation	MEF2 Reporter Positive Purkinje Cells (>4× Background) at t = 40 Min
Patched Cells
Vhold = −70 mV	2/20
Depolarization	3/20
Glutamate pulses	3/20
Depolarization + glutamate pulses	19/20
Vhold = −70mV (MEF2A+2D DKO)	1/20
Depolarization + glutamate pulses (MEF2A+2D DKO)	0/20
Non-patched Cells
No treatment	3/100
MEF2-VP16-ER, 16–26 h tamoxifen	100/100
MEF2-ΔDBD-VP16-ER, 16–26 h tamoxifen	2/100
Tamoxifen, 16–26 h	3/100
Phorbol-12,13-diacetate, 200 nM, 10 min	97/100
MEF2D S444A	2/100

A MEF2-activity reporter was created consisting of a triple repeat of the MEF2-response element (MRE) fused to a minimal promoter of the fos gene (that does not itself confer glutamate or depolarization responsiveness; Flavell et al., 2006) inserted in front of destabilized EGFP. Using a gene gun, this reporter plasmid was delivered to Purkinje cells, together with a separate dsRedexpress2 plasmid as a marker, 3 days after the culture was prepared. Measurements were then made 7–10 days after transfection.