Chlamydia trachomatis Infection Protocol

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)).

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Pedrosa A.T., Murphy K.N., Nogueira A.T., Brinkworth A.J., Thwaites T.R., Aaron J., Chew T.L, & Carabeo R.A. (2020). A post-invasion role for Chlamydia type III effector TarP in modulating the dynamics and organization of host cell focal adhesions. The Journal of Biological Chemistry, 295(43), 14763-14779.

Publication 2020

COS7 HeLa 229 DMEM Gastrografin

C trachomatis Cells Density gradient centrifugation Diagnostics Fetal bovine serum Gastrografin Gentamicin Glutamine Hela Nih3t3

Corresponding Organization : University of Nebraska Medical Center

Other organizations : University of Aberdeen, Washington State University, Janelia Research Campus, Imaging Center, Howard Hughes Medical Institute

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Variable analysis

independent variables

Cell lines used: COS7, NIH3T3, HeLa 229, MEFs vcl-/- and matched MEFs vcl+/+

dependent variables

Not explicitly mentioned

control variables

Cell culture media composition: Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum, 2 mM L-glutamine, and 10 μg/ml gentamicin
C. trachomatis serovar L2 (L2/434/Bu) used as the infectious agent

positive controls

None specified

negative controls

None specified

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