Supplemental Materials
This article contains the following supporting material:
- Supplemental Materials
- Movie 6 - Figure 6. mov 3-6. cAMP-induced redistribution of STIM2 and GluA1 in hippocampal neurons.Movie 3: TIRF imaging of YFP-STIM2 in the soma and proximal dendrites of a hippocampal neuron (DIV 7) following forsk/rolipr treatment. Movies 4 and 5: Dual-color TIRF imaging of mCherry-STIM2 (movie 4) and SEP-GluA1 (movie 5) in a hippocampal neuron (DIV7) following forsk/rolipr treatment. Movie 6: Dual color confocal imaging of YFP-STIM2 (green) and mCherry (magenta) upon forsk/rolipr addition, in a dendritic segment of a DIV 19 neuron. Frame at which forsk/rolipr is added is marked in all movies. Time is in min and sec. Scale bar: 5 μm.
- Movie 1 - Figure 4. mov 1 and 2. FRET imaging of PKA activity in control and STIM2-silenced cells.Time-lapse AKAR3EV FRET imaging of hippocampal neurons (DIV21) co-expressing AKAR3EV, mCherry and scramble (movie 1) or STIM2 (movie 2) shRNAs. A dendritic segment is shown. AKAR3EV FRET is displayed as the FRET/CFP ratio, and is pseudocolored. Forsk/rolipr was added at ~0.5 sec into the movie. Movie played at 20 f/sec and sped up 600x.
- Movie 4 - Figure 6. mov 3-6. cAMP-induced redistribution of STIM2 and GluA1 in hippocampal neurons.Movie 3: TIRF imaging of YFP-STIM2 in the soma and proximal dendrites of a hippocampal neuron (DIV 7) following forsk/rolipr treatment. Movies 4 and 5: Dual-color TIRF imaging of mCherry-STIM2 (movie 4) and SEP-GluA1 (movie 5) in a hippocampal neuron (DIV7) following forsk/rolipr treatment. Movie 6: Dual color confocal imaging of YFP-STIM2 (green) and mCherry (magenta) upon forsk/rolipr addition, in a dendritic segment of a DIV 19 neuron. Frame at which forsk/rolipr is added is marked in all movies. Time is in min and sec. Scale bar: 5 μm.
- Movie 10 - Figure 8. mov 7-10. STIM2 is required for PKA-dependent surface delivery of SEP-GluA1 and spine enlargement.Dual color imaging of SEP-GluA1 (movies 7 and 9) and mCherry (movies 8 and 10) in hippocampal neurons (DIV 18) co-expressing scrambled (movies 7 and 8) or STIM2 (movies 9 and 10) shRNAs. Time at which forsk/rolipr is added is marked. Time is in min and sec. SEP-GluA1 intensity is color coded, according to the scale shown in Figure 7A. Scale bar, 5 μm.
- Movie 3 - Figure 6. mov 3-6. cAMP-induced redistribution of STIM2 and GluA1 in hippocampal neurons.Movie 3: TIRF imaging of YFP-STIM2 in the soma and proximal dendrites of a hippocampal neuron (DIV 7) following forsk/rolipr treatment. Movies 4 and 5: Dual-color TIRF imaging of mCherry-STIM2 (movie 4) and SEP-GluA1 (movie 5) in a hippocampal neuron (DIV7) following forsk/rolipr treatment. Movie 6: Dual color confocal imaging of YFP-STIM2 (green) and mCherry (magenta) upon forsk/rolipr addition, in a dendritic segment of a DIV 19 neuron. Frame at which forsk/rolipr is added is marked in all movies. Time is in min and sec. Scale bar: 5 μm.
- Movie 7 - Figure 8. mov 7-10. STIM2 is required for PKA-dependent surface delivery of SEP-GluA1 and spine enlargement.Dual color imaging of SEP-GluA1 (movies 7 and 9) and mCherry (movies 8 and 10) in hippocampal neurons (DIV 18) co-expressing scrambled (movies 7 and 8) or STIM2 (movies 9 and 10) shRNAs. Time at which forsk/rolipr is added is marked. Time is in min and sec. SEP-GluA1 intensity is color coded, according to the scale shown in Figure 7A. Scale bar, 5 μm.
- Movie 8 - Figure 8. mov 7-10. STIM2 is required for PKA-dependent surface delivery of SEP-GluA1 and spine enlargement.Dual color imaging of SEP-GluA1 (movies 7 and 9) and mCherry (movies 8 and 10) in hippocampal neurons (DIV 18) co-expressing scrambled (movies 7 and 8) or STIM2 (movies 9 and 10) shRNAs. Time at which forsk/rolipr is added is marked. Time is in min and sec. SEP-GluA1 intensity is color coded, according to the scale shown in Figure 7A. Scale bar, 5 μm.
- Movie 10 - Figure 8. mov 7-10. STIM2 is required for PKA-dependent surface delivery of SEP-GluA1 and spine enlargement.Dual color imaging of SEP-GluA1 (movies 7 and 9) and mCherry (movies 8 and 10) in hippocampal neurons (DIV 18) co-expressing scrambled (movies 7 and 8) or STIM2 (movies 9 and 10) shRNAs. Time at which forsk/rolipr is added is marked. Time is in min and sec. SEP-GluA1 intensity is color coded, according to the scale shown in Figure 7A. Scale bar, 5 μm.
- Movie 2 - Figure 4. mov 1 and 2. FRET imaging of PKA activity in control and STIM2-silenced cells.Time-lapse AKAR3EV FRET imaging of hippocampal neurons (DIV21) co-expressing AKAR3EV, mCherry and scramble (movie 1) or STIM2 (movie 2) shRNAs. A dendritic segment is shown. AKAR3EV FRET is displayed as the FRET/CFP ratio, and is pseudocolored. Forsk/rolipr was added at ~0.5 sec into the movie. Movie played at 20 f/sec and sped up 600x.
- Movie 5 - Figure 6. mov 3-6. cAMP-induced redistribution of STIM2 and GluA1 in hippocampal neurons.Movie 3: TIRF imaging of YFP-STIM2 in the soma and proximal dendrites of a hippocampal neuron (DIV 7) following forsk/rolipr treatment. Movies 4 and 5: Dual-color TIRF imaging of mCherry-STIM2 (movie 4) and SEP-GluA1 (movie 5) in a hippocampal neuron (DIV7) following forsk/rolipr treatment. Movie 6: Dual color confocal imaging of YFP-STIM2 (green) and mCherry (magenta) upon forsk/rolipr addition, in a dendritic segment of a DIV 19 neuron. Frame at which forsk/rolipr is added is marked in all movies. Time is in min and sec. Scale bar: 5 μm.