Fig. 2: Ca elevation opens compartments that extend deeply into the cytoplasm.
From: TMEM16F and dynamins control expansive plasma membrane reservoirs
A Reversible fluorescent PM staining by TB before and after Ca elevation in WT and TMEM16F-null Jurkat cells reveal TB fluorescence as a relatively uniform indicator for PM changes. Changes of TB fluorescence (%) are plotted against changes of Cm (%) in response to Ca elevation. Linear regression of results with 95% C.I. B Representative fluorescence line scans during TB application in Jurkat WT and TMEM16F-null cells before (blue) and after (red) Ca elevations. Quantification of average PM fluorescence depth in WT (n = 4) and TMEM16F-null Jurkat cells (5), TMEM16F-null HEK cells with (4) and without (6) rescued TMEM16F expression and BHK cells with (5) and without spermine (6). Confocal imaging of a BHK cell during TB application and after washout, before and after Ca elevation via NCX1. Extracellular continuity of labeled compartments (3) is indicated by washout (4) D As in (C) Trehalose exposure at low ionic strength reverses PM expansion (4), n = 11. All data were analyzed from the total number of independent cells (n) from a minimum of two experimental repeats and expressed as mean ± s.e.m. Paired Student’s t-test was used for comparing two groups for (B) and (D). All Scale bars: 5 µm.
