P. gingivalis, but not S. gordonii or F. nucleatum, degrades JAM1 in IHGE cells.
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(A) Schematic view of the structure of Myc-mCherry–tagged HA-inserted JAM1. The immature form of Myc-mCherry–tagged HA-inserted JAM1 was used as an internal control to monitor degradation flux of the mature form of HA-inserted JAM1. IHGE cells were transiently transfected with Myc-mCherry–tagged HA-inserted JAM1 plasmid. Following 48 h of incubation, cells were infected for 1 or 2 h with the indicated bacteria at an MOI of 100. The cells were then analyzed by immunoblotting using the indicated antibodies. (B–E) IHGE cells were transiently transfected with the HA-inserted JAM1 plasmid. Following 48 h of incubation, the cells were infected with P. gingivalis ATCC 33277 (B), P. gingivalis TDC60 (C), S. gordonii DL-1 (D), or F. nucleatum ATCC 25586 (E) at an MOI of 100 for the indicated times. The cells were then analyzed by immunoblotting with the indicated antibodies.