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. 2018 Apr 30;37(11):e97943. doi: 10.15252/embj.201797943

Figure 1. Ago2 association with GW182 in neuronal dendrites increases in response to NMDAR stimulation.

Figure 1

  1. Endogenous Ago2‐GW182 and Ago2‐DDX6 interactions increase in response to NMDAR stimulation. Cortical neuronal cultures were exposed to NMDA or vehicle for 3 min, and lysates were prepared 10 min after NMDA washout and immunoprecipitated with Ago2 antibodies. Proteins were detected by Western blotting. Graph shows quantification of Ago2‐GW182 interaction, normalised to vehicle control; n = 5. *P < 0.05; ***P < 0.001; t‐test; mean ± SEM.
  2. Analysis of endogenous Ago2‐GW182 co‐localisation in cortical neuronal cultures. Cortical neuronal cultures were exposed to NMDA or vehicle for 3 min, fixed 10 min after NMDA washout, permeabilised and co‐stained with Ago2 and GW182 antibodies. Representative whole‐cell images are shown. Scale bar = 50 μm.
  3. Endogenous GW182‐Ago2 co‐localisation increases in response to NMDAR stimulation in neuronal dendrites. Images show dendrites taken from boxed region in (B), above. Graph shows Pearson's co‐localisation coefficients; n = 4 independent experiments (18–24 cells per condition). *P < 0.05, t‐test. Scale bar = 10 μm. Mean ± SEM.
  4. Line‐scan analyses of Ago2 and GW182 fluorescence intensities in control and NMDA‐stimulated dendrites shown in (C).
  5. NMDAR stimulation has no effect on endogenous Ago2‐GW182 co‐localisation in neuronal cell bodies. Images show cell bodies taken from boxed region in (B). Graph shows Pearson's co‐localisation coefficients; n = 4 independent experiments (18–20 cells per condition), t‐test. Scale bar = 10 μm. Mean ± SEM.

Source data are available online for this figure.