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Data were compared with those of the INS-GFP− fraction from the same cultures, as well as from fetal pancreas, adult pancreas and adult islets.
We transiently expressed YFP-Drp1 in the same cell lines and then imaged mitochondria to detect YFP, using non-transfected (YFP negative) cells in the same cultures as controls (Fig. 6A D).
For viability tests using exclusion of fluorescent dyes as an indicator of metabolic activity, aliquots of the same cultures as above were stained with either propidium iodide or phloxine B. For propidium iodide staining, cells were centrifuged, washed twice with PBS, and incubated with 3 µg/ml of the dye for 40 min on ice in darkness.
TEER and permeability measurements can be performed with the same cultures as used for PMN transmigration.
IP-10 (A ), TNF (B ), IL-17 (C ), VEGF (D ), IL1-β (E ) and G-CSF (F ) production in same cultures as described in Figure S1. Figure S3.
Exflagellation was determined in the same cultures as described above at different time points (10 22 min) after activation of gamete formation.
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