Ultrapure P. gingivalis LPS (UP Pg LPS) (Invivogen, cat: tlrl-ppglps, lot: PPG-3701 and 3801), Standard P. gingivalis LPS (STD Pg LPS) (cat: tlrl-pglps, lot: LPG 37-01 and 38-01) were from InvivoGen (France). The STD Pg LPS were obtained by classical methods using hot-phenol extraction using the bacterial strain ATCC 33227 and UP Pg LPS preparations by enzymatic removal of lipoprotein from STD Pg LPS (Invivogen). Ultrapure lipopolysaccharide from E. coli O111:B4 (UP Ec LPS) (cat: tlrl-3pelps, lot: L3P 37-02) were from InvivoGen (France).
RAW264.7-Blue (RAW-Blue), HEK-Blue hTLR2-hCD14 (HEK-Blue hTLR2), HEK-Blue Null, HEK-Blue hTLR4-hCD14-hMD2 (HEK-Blue hTLR4), and HEK-Blue mTLR4-mCD14-mMD2 (HEK-Blue mTLR4) were from InvivoGen (France). BV2 was obtained as described previously35 (link). J774.1 (TIB-67), 3T3-L1 (CL-173), bEnd.3 (CRL-2299), and RAW264.7 (TIB-71) were from ATCC (USA), and all of these cells line express functional TLR2 and TLR4. TNF-α was from R&D systems (USA). Except when indicated, all other reagents were from Sigma-Aldrich (France). NFkB activity was performed using Quanti-blue assay following the manufacturer’s protocols (InvivoGen).
RAW264.7-Blue (RAW-Blue), HEK-Blue hTLR2-hCD14 (HEK-Blue hTLR2), HEK-Blue Null, HEK-Blue hTLR4-hCD14-hMD2 (HEK-Blue hTLR4), and HEK-Blue mTLR4-mCD14-mMD2 (HEK-Blue mTLR4) were from InvivoGen (France). BV2 was obtained as described previously35 (link). J774.1 (TIB-67), 3T3-L1 (CL-173), bEnd.3 (CRL-2299), and RAW264.7 (TIB-71) were from ATCC (USA), and all of these cells line express functional TLR2 and TLR4. TNF-α was from R&D systems (USA). Except when indicated, all other reagents were from Sigma-Aldrich (France). NFkB activity was performed using Quanti-blue assay following the manufacturer’s protocols (InvivoGen).
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