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. 2019 Sep 24;5(11):1768–1776. doi: 10.1021/acscentsci.9b00567

Figure 4.

Figure 4

Multiplexed PPI analysis with split BS2. (a) Chemilum-CM is unmasked by esterase activity and generates a photon. (b) Vector system to monitor extracellular PPIs. (c) HEK293T cells were cotransfected with plasmids shown in part b. Rapamycin (blue) or a DMSO control (gray) were added to cells for 0–6 h. Media was replaced with Chemilum-CM and analyzed for luminescence. HEK293T control cells (white) were similarly analyzed. (d) HEK293T cells cotransfected as in part c or HEK293T control cells (triangle) were incubated with rapamycin (blue) or a DMSO control (gray). After 6 h, the cells were analyzed with Chemilum-CM as in part c. (e) Vector system to simultaneously monitor extracellular and intracellular PPIs. (f) HEK293T cells were transfected with all four plasmids shown in part d. Rapamycin or a DMSO control was added to the cells for 24 h. Media was replaced with Chemilum-CM (10 μM) and analyzed for luminescence (blue). The cells were then rinsed, incubated with furimazine, and analyzed for bioluminescence (orange). Error bars are the standard deviation for n = 4 replicates. Unpaired t test; *P < 0.01, **P < 0.001, ***P < 0.0001.