<p>(<b>A</b>) Schema of 2-NBDG accumulation and retention (glycogen labeling) experiments. (<b>B</b>) 2-hour 2-NBDG accumulation in the presence of 10 mM D-glucose. Upper panel: green fluorescence intensity (Fluor) images from 2-NBDG alone. These images were obtained (immediately after replacing with fresh mTeSR1 medium) by non-saturated time-exposure guided by an autoexposure software (Zeiss Inc.). Lower panel: the corresponding phase images of the upper panel. Only brightness was adjusted in phase images (presented in both B and C) to enhance the image presentation in this figure. (<b>C</b>) 2-NBDG retention and glycogen labeling carried out in the presence of 10 mM D-glucose and absence of 2-NBDG. Upper panel: unique fluorescence loci (dots) were derived from 2-NBDG signals as detailed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0142554#pone.0142554.g005" target="_blank">Fig 5</a>. (<b>D</b>) Quantitative analysis of mean fluorescence intensity (FI) in Fig 6B. (<b>E</b>, <b>F</b>) Quantitative analysis of 2-NBDG retention and glycogen labeling by measuring mean fluorescence intensity (FI, arbitrary units) from at least 4 random colonies (E) and by counting 2-NBDG loci (F). Columns represent mean fluorescence intensity measured from at least 4 random colonies and bar standard deviations. Scale bars represent 100 μm.</p
