Goat anti mouse hrp conjugated secondary antibodies

Cells were treated with the OMV or left untreated for the indicated times. Then cells were washed twice with cold PBS, lysed in NuPAGE LDS sample buffer with reducing agent (Invitrogen) and sonicated. Cell extracts were boiled for 5min at 95°C, separated by SDS-PAGE and transferred to PVDF membranes (Invitrogen). The membranes were blocked in 5% BSA in 0.1% Tween-20 in TBS and probed with primary antibodies. The primary antibodies used were Rabbit anti-LC3 (Cell Signaling S 3868), Mouse anti-actin (Abcam Ab 3280). After incubation with Goat anti-Rabbit and Goat anti-Mouse HRPconjugated secondary antibodies (Promega), proteins were visualized using chemiluminescent peroxidase substrat detection reagents (Sigma), and acquired by the ChemiDoc XRS+ System (BioRad). Densitometric analyses of immunoblots were performed using the ImageJ software.