Hela 229

The human HeLa cell line [HeLa 229 (ATCC^® CCL-2.1^TM)] used in this study was obtained from ATCC (ATCC, Manassas, VA, USA). The cells were cultured in complete medium, which is Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Thermo Fisher Scientific, Waltham, MA, USA), supplemented with 10% fetal bovine serum (Merck KGaA, Darmstadt, Germany) and antibiotics (100 U/mL penicillin and 100 μg/mL streptomycin) (Gibco, Thermo Fisher Scientific, Waltham, MA, USA), and maintained under a humidified atmosphere of 37 °C, 5% CO₂. The cells were sub-cultured every 2–3 days.

COS7 (ATCC CRL-1651), NIH3T3 (kindly supplied by Hector Aguilar-Carreño ATCC CRL-1658), and HeLa 229 (ATCC CCL-2.1) were used in this paper. MEFs vcl^−/− and matched MEFs vcl^+/+ (59 (link)) were kindly provided by Dr. Wolfgang Ziegler (Hannover Medical School). Cells were cultured using Dulbecco's modified Eagle's medium (DMEM) (Thermo Fisher Scientific, 11960-085). Media were supplemented with 10% fetal bovine serum (Sigma, F0804-500ML), 2 mm l-glutamine, and 10 μg/ml gentamicin. C. trachomatis serovar L2 (L2/434/Bu) was propagated in HeLa 229. EBs were harvested by discontinuous density gradient centrifugation in Gastrografin (Bracco Diagnostics), as described previously (16 (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 ).

Seven CCa cell lines, including Siha, HeLa, Caski, MS751, ME180, C33A and HeLa229, and normal cervix derived cell line H8 were purchased from ATCC and cultured in a humidified atmosphere with 5% CO₂ at 37 °C. Human lymphatic endothelial cells (HLECs) were obtained from ScienCell Research Laboratories and maintained in the recommended endothelial cell medium (ScienCell, CA). All cell lines were cultured in complete medium with 10% FBS (Gibco, USA) as previously described^{31 (link)}.

VERO (ATCC CCL-81), HeLa 229 (ATCC CCL-2.1), NIH/3T3 (ATCC CRL-1658), A431 (ATCC CRL-1555), HS578T (ATCC HTB-126), HT-3(ATCC HTB-32), MDA-MB-231 (ATCC HTB-26), SKOV3 (ATCC HTB-77), and PC-3 (ATCC CRL-1435) were originally from ATCC. HEK 293FT was from Thermo Fisher Scientific (cat. no. R70007). Mouse embryonic fibroblasts (MEFs) were generated from embryos of SAMD9L^−/− and SAMD9L^+/+ mice according to standard protocols. Human foreskin fibroblasts (HFFs), described in [33 (link)], were kindly provided by Dr. Zhilong Yang.

Human CCa cell lines (SiHa, HeLa, ME180, C33A, Caski, HeLa229, and MS751) and a normal cervical cell line (H8) were purchased from the American Type Culture Collection (ATCC, USA) and ZQXZBIO (Shanghai Zhong Qiao Xin Zhou Biotechnology Co., Ltd.). Siha, HeLa, and H8 cells were cultured in DMEM (Gibco, China), MS751 and C33A cells were cultured in MEM (Gibco, USA), while HeLa229 and Caski cells were cultured in RPMI1640 (Gibco, USA). All media was supplemented with 10% fetal bovine serum (FBS) (Gibco, USA) and 1% penicillin/streptomycin (Gibco, China). Cells were cultured in a humid atmosphere with 5% CO₂ at 37 °C. In 2018, all of the cell lines used were tested for authenticity by short tandem repeat (STR) genotyping; the cell lines were also screened for mycoplasma contamination (e-Myco Mycoplasma PCR Detection Kit; iNtRON).