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Figure 2 Pre-Treatment of MSCs With GFs Enhanced the Formation of Functioning Gap Junction With CMCs

(A and B) The MSCs labeled with 1.1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) (red) and calcein-AM (green) were cocultured with unlabeled CMCs. Only green calcein-AM is transmittable through functioning gap junction. Cells having green fluorescence without red denote CMCs that are connected with labeled MSCs through functioning gap junction. These cells were scarcely observed in the coculture with untreated MSCs (A), whereas they were frequently observed in the coculture with GF-treated MSCs (B). (C) After addition of heptanol into coculture condition with treated MSC and CMCs, there are no CMCs having green fluorescence alone, corroborating that calcein transfer was mediated through gap junction (magnification x200). (D) Treatment with GFs significantly increased intercellular communication between MSCs and CMCs. Data were derived from 5 randomly chosen microscopic fields of 3 separate experiments. HPF = high power field; other abbreviations as in Figure 1.