Supplementary Figure 7: OPHN1 fluorescence intensity was positively correlated with p-eIF2α levels at individual synapses of cultured hippocampal neurons.
From: Translational control of mGluR-dependent long-term depression and object-place learning by eIF2α
a-d) Representative two-channel immunostaining for p-eIF2α and OPHN1 from a dendritic segment in 14DIV control- (b), DHPG- (c), and Sal003-treated (d) hippocampal neuron cultures fixed 15 min after treatment onset. Framed areas (white squares) in (b) are expanded on the left (a). p-eIF2α and OPHN1 fluorescence intensity (FI) were positively correlated at individual synapses. b-d) As shown in the scatter plots depicting data from a total of 315 synapses from 9 neurons analyzed per condition. Lines in b-d represent the regression lines of the log-transformed population FI data (in AU). Slopes (m) and correlation coefficients (r) of the respective regression lines are indicated. Scale bars indicate 2mm. e-f) Bar graphs of the average FI of synaptic p-eIF2α (e) and OPHN1 (f) normalized to vehicle-treated controls [p-eIF2a: control = 1.0 ± 0.06, DHPG = 2.2 ± 0.1, Sal003 = 2.4 ± 0.07, F(2,24)=19.6, p=9X10-6; OPHN1: control = 1.0 ± 0.05, DHPG = 1.9 ± 0.08, Sal003 = 1.8 ±0.1, F(2,24)=21.0, p=5.4X10-6]. Statistical significance was assessed by one-way ANOVA. Results are displayed as mean ± SEM.
