Transcript emboj7601969-sup

A
116 kDa
97 kDa
116 kDa
97 kDa
Blot: anti GluR1
111 kDa
Blot: anti GluR2L
111 kDa
Blot: anti GluR2L
Blot: anti GluR4
111 kDa
Blot: anti GluR2 (N-term)
Blot: anti GluR2/3
116 kDa
97 kDa
B
Blot GluR2L
Blot GluR2L
Blot GluR4
Blot GluR4
Supplementary Figure 1: Specificity of AMPA-R antibodies used in this study. GluR2L
and GluR4 are specifically immunoprecipitated by their respective antibodies from
cultured neurons and rat brain. A: HEK293T cells were transfected with cDNA constructs
coding for full-length GluR1, GluR2L or GluR4 (left panels), or with GluR2L, GluR2 short
form or empty vector (pRK5) (right panels). Lysates were immunoblotted with the indicated
antibodies. B: Cortical cultured neuronal lysates (DIV17-21, left panels) or rat brain P2
membrane fractions (right panels) were prepared as in Methods and subjected to
immunoprecipitation and Western blotting with the indicated antibodies. These data show that
these receptors can be specifically immunoprecipitated and blotted by their cognate antibodies
from both cultured neurons and rat brain. In addition, these results suggest that GluR2L and
GluR4 do not heteromultimerize, although it is unclear to what degree receptors containing
these two AMPA-R subunits are restricted to different neuronal cell types, or are present at
different locations within the same neuron.