abcC8⁻ cells have streaming defects and have impaired cAMP release from EVs. (A) Bright-field mages of WT and abcC8⁻ cells plated on DB agar taken 4, 5, and 6 h after the initiation of starvation. Images representative of three independent experiments. (B) Bright-field images of micropipette streaming assay showing the response of WT and abcC8⁻ cells to a micropipette filled with 1 µM cAMP. Images were taken 20 min after the micropipette was added. Images representative of three independent experiments. (C) Bright-field images of micropipette streaming assay showing the response of WT, abcC8⁻, and ABCC8-RFP/abcC8⁻ cells to a micropipette filled with 1 µM cAMP. Images were taken 20 min after the micropipette was added. Images representative of three independent experiments. (D) Western blots of ABCC8-RFP expression in various preparations, as depicted. Results representative of three independent experiments. (E) Representative confocal fluorescent maximum-intensity projection image overlaid on bright-field image of differentiated ABCC8-RFP/abcc8⁻ cells streaming toward an aggregate. (F and G) Time-dependent release of cAMP from intact (F) or lysed (G) EVs isolated from WT and abcC8⁻ cells. Results presented as mean ± SEM of three independent experiments.