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. Author manuscript; available in PMC: 2015 Sep 1.
Published in final edited form as: Biomaterials. 2014 Jun 24;35(29):8348–8356. doi: 10.1016/j.biomaterials.2014.06.018

Figure 4. The ISO-induced PKA activation and receptor internalization form a feedback loop.

Figure 4

(A) Time courses of emission ratio in the cells pretreated with H89 (20 μM), a PKA inhibitor, in response to ISO. H89 inhibits FRET emission ratio in ECFP-AKAR2-YPet transfected HMSCs. Similar results were observed in three independent experiments. (B) H89 inhibits ISO-induced internalization of β2-AR on both hard (40 kPa) and soft (0.1 kPa) gels. Similar results were observed in three independent experiments. (C) MβCD treatment inhibits ISO-induced β2-AR internalization in HMSCs cultured on both hard (40 kPa) and soft (0.1 kPa) gels. (D-E) FRET/ECFP emission ratio images and time courses revealed that the disruption of lipid rafts and receptor internalization by MβCD blocks ISO-induced PKA activation on hard gels (40 kPa) (n=6, ***P<0.001), whereas untreated cells still show the FRET increase in response to ISO (n=7).