Hyperthermia Differently Affects Connexin43 Expression and Gap Junction Permeability in Skeletal Myoblasts and HeLa Cells
Figure 4
Measurement of Cx43-EGFP GJ channel number in JPs and hemichannel density in the plasma membrane of HeLa cells. (a) To estimate the total fluorescence of Cx43-EGFP JP (green), emitted light was integrated over the ROI encircling JP. (b) To estimate fluorescence intensity of the single Cx43-EGFP GJ channel, emitted light was integrated over the ROI situated in the horizontal JP of overlapping cells. (c) TIRF microscopy was used to estimate the density of Cx43-EGFP hemichannels (red) on the cell surface. Emitted light was integrated over the ROI situated in the site with most homogenous fluorescence. (d) To estimate the fluorescence intensity of the single Cx43-EGFP GJ hemichannel, emitted light was integrated by a TIRF microscope over the ROI situated in the horizontal JP of overlapping cells. Details are presented in the main text. Scale bar 10 μm.
