Data Set 1: Live cell microscopy videos of cellular distribution of PTEN:EGFP and PTEN-L:EGFP after 50µM glutamate stress in mouse primary neurons (Fig. 3a-b)

This dataset contains 8 live cell microscopy timelapse video files in <b>.avi</b> format. The imaging targets murine primary neurons using enhanced green fluorescent protein (EGFP).<br>The related study investigates the function and regulation of PTEN-L in neurons in the context of brain ischemia. Exogenous, EGFP-tagged PTEN-L or the shorter variant PTEN were expressed in primary neurons after knock out of endogenous PTEN. Oxygen-glucose deprivation or exposure to 50 µM glutamate were applied to neurons to test their response to ischemic-like stress.<br><b>Methods</b><br>Cells expressing PTEN:EGFP or PTEN-L:EGFP were imaged every 10 min for 90 min. The time-lapse movies compiled from the 90 min observation. Primary neurons derived from conditional PTEN knockout mice were transduced with both <i>Cre</i>-delivering lentiviral particles (LVPs) or control LVPs and <i>Pten</i>-delivering LVPs or control LVPs as described above and seeded in 8-well microscopy dishes (Ibidi). DIV 9 primary neurons were imaged while maintaining culturing conditions (20 % oxygen, 5 % CO₂ and 37°C). A confocal microscope (Nikon Ti2) with uniform spinning disk illumination (Andor Borealis), an EMCCD Camera (iXon3 DU-888 Ultra) and 60x Plan Apo oil objective (Nikon) was used to acquire images. 8 positions across 4 wells were selected and images of z-planes (30 - 40 µM with 1 µM intervals) were taken with a laser exciting at 488 nm (&gt;8 mW; 20 %) detecting EGFP and a laser exciting at 561 nm (&gt;15 mW; 9 %) detecting mCherry. Next, cells were treated with 50 µM glutamate and each position was imaged every 10 min for 90 min using above described settings. Fiji software was used to compile time lapse videos over the 90 min observation period. <br>