Anti ha 11 clone 16b12 mab

Anti-HA.11 clone 16B12 mAb was purchased from Covance; anti-FAK (phospho-Tyr-397 (pY397-FAK)) pAb and anti-vinculin mAb were from Abcam, and anti-p34^Arc pAb was purchased from Millipore. Anti-rabbit or anti-mouse IgG secondary antibodies were either Alexa Fluor 488 or 594 were purchased from Invitrogen. Phalloidin conjugated to Alexa Fluor dye was purchased from Invitrogen. Cos7 (ATCC CRL-1651) and HeLa 229 (ATCC CCL-2.1) were routinely grown in DMEM supplemented with 10% FBS, 2 mml-glutamine, and 10 μg/ml gentamicin. Subcultivation was at a 1:4 ratio. The cells were used at passage <15. FAK^−/− mouse embryo fibroblasts (CRL-2644) and matched FAK^+/+ cells (CRL-2645), purchased from LGC standards, were cultured in DMEM + 10% FBS and subcultured at a 1:4 ratio. Cultured cells were grown in a humidified 5% CO₂ incubator at 37 °C. Chlamydia trachomatis serovar L2 and C. caviae strain guinea pig inclusion conjunctivitis (GPIC) were propagated in HeLa cells grown in DMEM + 10% FBS supplemented with gentamicin (10 μg/ml). Harvest of elementary bodies was by discontinuous density gradient centrifugation in Renografin (Bracco Diagnostics), as previously described (35 (link)).

Anti-HA.11 clone 16B12 mAb was purchased from Covance; anti-FAK (phospho Y397) pAb and anti-vinculin mAb were from Abcam. Anti-rabbit or anti-mouse IgG secondary antibodies, either Alexa Fluor 488 or 594 were purchased from Invitrogen. Phalloidin conjugated to Alexa Fluor dye was purchased from Invitrogen. Cos7 (ATCC CRL-1651) and HeLa 229 (ATCC CCL-2.1) were routinely grown in DMEM supplemented with 10% FBS, 2 mM L-glutamine, and 10 μg/ml gentamicin. Subcultivation was at 1:4 ratio. The cells were used at passage < 15. FAK^−∕− mouse embryo fibroblasts (MEFs; CRL-2644) and matched FAK^+∕+ cells (CRL-2645) were purchased from LGC standards. vcl^−∕− and matched vcl^+∕+ MEFs (Marg et al., 2010 (link)) were generously provided by Dr. Wolfgang Ziegler (Hannover Medial School). All MEFs were cultured in DMEM + 10% FBS and subcultured at 1:4 ratio. Cultured cells were grown in a humidified 5% CO2 incubator at 37°C. C. caviae strain GPIC was propagated in HeLa cells grown in DMEM + 10% FBS supplemented with gentamicin (10 μg/ml). Harvest of elementary bodies was by discontinuous density gradient centrifugation in Renografin (Bracco Diagnostics), as previously described (Caldwell et al., 1981 ).