Fig 2: P. gingivalis infection induces an increased expression of the TIR domain containing proteins in EC. Immunofluorescence staining of uninfected and infected EC with P. gingivalis (Pg) (MOI:100) for 24 h. In red: MAL, MyD88, SARM, TRAM, and TRIF; and in blue: DAPI (nuclear staining). GFP-EC were used in this experience. All images were acquired at 20× magnification.

Fig 3: P. gingivalis infection induces an increased expression of the TIR domain containing proteins in GEC. Immunofluorescence staining of uninfected and infected GEC with P. gingivalis (Pg) (MOI:100) for 24 h. In green: MAL, MyD88, SARM, TRAM, and TRIF; in red: phalloidin; and in blue: DAPI (nuclear staining). All images were acquired at 20× magnification.

Fig 4: Co-immunoprecipitated proteins using Dynabeads Protein G, identifying interactions between the five TIR domains containing proteins (MyD88, MAL, TRIF, TRAM, SARM) and TLR2 and TLR4 in GEC (A) and EC (B), by SDS-PAGE and blotting of the protein complexes. In each column, the primary precipitated protein by the Ab capture is indicated in the upper part. Then, in each band, the presence and expression of the different proteins that are interacting at the time of precipitation are indicated with the arrows.

Fig 5: P. gingivalis significantly increased mRNA expression of TIR domain containing proteins in EC. The mRNA expression of MAL and MyD88 with and without their TIR terminal domain, TLR2, TLR4, and other TIR domain containing proteins: TRIF, TRAM, and SARM in GEC infected with P. gingivalis (Pg) (MOI:100) and stimulated with Pg-LPS, 1µg/mL) for 24 h was measured by qRT-PCR. All data was expressed as the mean mRNA expression normalized with the ß-actin expression ± SD from three independent experiments *: P < 0.05 between cells infected or stimulated against control (unstimulated cells).