Sc 57840

For immunofluorescence as well as immunoblotting experiments the following antibodies were used. Rabbit monoclonal antibody (ab131209 from Abcam), Rabbit polyclonal antibody (ab113895 from Abcam) against human PI15, mouse monoclonal anti-flag antibody for immunoblotting (F3605, Sigma), mouse monoclonal anti-Flag antibody for immunostaining (F1804, Sigma), mouse monoclonal against human RFX5 (sc-271756, Santa Cruz Biotechnology), mouse monoclonal against human cytokeratin 8 (sc-58737, Santa Cruz Biotechnology), rabbit polyclonal against human vimentin (sc-5565, Santa Cruz Biotechnology), mouse monoclonal against chlamydial Hsp60 (sc-57840, Santa Cruz Biotechnology), mouse monoclonal against human beta-actin (A5441, Sigma). Antibodies against CPAF, cHtrA, IncA were kindly provided by Guangming Zhong and George Haecker. For immunoprecipitation experiments, anti-Flag M2 antibody bound to magnetic beads (Sigma) were used.

Treated cells were incubated for 0, 12, 24, 36, or 48 h, then the cellular proteins were lysed by RIPA (Invitrogen, 89900) supplemented with a protease and phosphatase inhibitor cocktail (Invitrogen, 78440), and incubated with SDS-PAGE loading buffer (Reducing) (Cwbio, CW0027) at 100°C for 10 min. Antibodies used for western blotting were as follows: anti-RSK1 p90 (phospho T359 + S363) antibody (1:1,000, ab32413, Abcam), anti-RSK1 p90 antibody (1:1,000, ab32114, Abcam), anti-Phospho-p44/42 MAPK (Erk1/2) (1:1,000, 4370S, Cell signaling), anti- p44/42 MAPK (Erk1/2) (1:1,000, 4695S, Cell signaling), anti-cHSP60 (1:2,000, sc-57840, Santa Cruz), anti-GAPDH (1:1,0000, ab181602, Abcam), anti-rabbit IgG-HRP-linked antibody (1:5,000, 7074S, Cell signaling), and anti-mouse IgG-HRP-linked antibody (1:5,000, 7076S, Cell signaling). Blots were imaged on a ChemiDoc MP Imaging System (Bio-Rad).