Regulation of Hemichannels and Gap Junction Channels by Cytokines in Antigen-Presenting Cells
Figure 4
IFN-γ or IL-6 increases dye uptake in dendritic cells. Bone-marrow-derived dendritic cells (BMDCs) from balb/c mice were differentiated with 40 ng/mL GM-CSF and IL-4 for 7 days. At day 7, BMDCs were treated for 6 h with IFN-γ (10 ng/mL), IL-6 (10 ng/mL), or both, and ethidium uptake was evaluated in time-lapse experiments (Olympus BX 51W1I). (a) Left: fluorescence images of ethidium after 9 min of uptake. Scale bar: 50 μm. Right: ImageJ surface plot analysis of fluorescence intensity of the region indicated in the field (dotted square). (b) Top: time-lapse ethidium uptake under control conditions (white circles) or after 6 h treatment with IL-6 (yellow triangles) or IFN-γ (blue diamonds). Each point corresponds to the mean of 30 cells. After 10 min of recording, La3+ (200 μM) was added to the bath solution to block connexin hemichannels. Bottom: graph showing the basal dye uptake rate and the effect of La3+ on BMDCs after treatment with IFN-γ (blue bars), IL-6 (yellow bars), or both (green bars). Each bar corresponds to the mean ± SE (% of control condition, dotted line) of 3 independent experiments.
