Journal: Oncotarget

Article Title: Tumor suppressor REIC/DKK-3 and co-chaperone SGTA: Their interaction and roles in the androgen sensitivity

doi: 10.18632/oncotarget.6488

Figure Lengend Snippet: A. The yeast two-hybrid analysis was conducted using pPC86 (AD)/full-length human SGTA (derived from a normal heart cDNA library) and pDBLeu (BD)/full-length human REIC/DKK-3 plasmids. The blue colonies indicate those with an interaction between the two proteins. B. For the pull-down (PD) assay, the full-length cDNA of human REIC/DKK-3 and SGTA was cloned into the pFN21A and pMACS Kk.HA-C plasmids, respectively. Cell lysates from Halo-tagged REIC/DKK-3- and/or HA-tagged SGTA-transfected 293T cells were analyzed. The sample pulled down using Halo-tagged REIC/DKK-3 was analyzed by Western blotting (WB) using anti-HA antibody. C. REIC/DKK-3 and SGTA protein expression in 293T, PC3 and Hs68 cells was analyzed by Western blotting. Coomassie Brilliant Blue (CBB) staining of the membrane is shown as a loading control. D. The co-localization of REIC/DKK-3 and SGTA was examined by double immunofluorescence staining and observed by fluorescence microscopy. The images in green and red show the intracellular localization of REIC/DKK-3 and SGTA, respectively. The areas of overlap between REIC/DKK-3 and SGTA are shown in yellow in the merged image.

Article Snippet: The normal human foreskin fibroblast cell line Hs68 was provided by JCRB Cell Bank (Osaka, Japan).

Techniques: Derivative Assay, cDNA Library Assay, Clone Assay, Transfection, Western Blot, Expressing, Staining, Membrane, Control, Double Immunofluorescence Staining, Fluorescence, Microscopy

Journal: BMC Microbiology

Article Title: First human cell-based cultivation system for the syphilis spirochete Treponema pallidum

doi: 10.1186/s12866-026-04856-5

Figure Lengend Snippet: Human foreskin fibroblast cell lines support the growth of T. pallidum in vitro . A Human foreskin fibroblast cell lines (MoNa, HFF1, and HFFC) support the growth of T. pallidum in long-term, continuous, in vitro cultivation. While cultivation of strain SS14 was discontinued after 10 weeks, in vitro cultures of the DAL-1 strain have been ongoing for over one year. During this period, scheme of subcultures was optimized (up to 1:20). A booster passage (*), extending the standard 7-day subculture to a 14-day period with a cultivation medium exchange on day 7, was employed to enrich the treponemal culture during critical declines. In vitro cultivation was performed in triplicate (i.e., in three cultivation wells) and the mean values for each subculture are presented in graph. Source data from individual wells are shown in Supplementary Table . B Parallel cultivation for seven days ( n = 15) with defined DAL-1 inoculum (10 6 treponemes) validated the differences in growth support of individual foreskin cell lines. Treponemal growth on human foreskin fibroblast cells was slower compared to growth on rabbit epithelial Sf1Ep cells. Red bar, mean. C Human foreskin-based cultivation system supports the growth of various T. pallidum strains ( n = 8), from the Nichols-like as well as the SS14-like cluster. Note that human foreskin fibroblast cells were prepared as an equal mixture of three tested cell lines. Each T. pallidum strain was cultivated in a single in vitro well, representing a sole experimental replicate used for data acquisition

Article Snippet: Human foreskin fibroblasts HFF1 (SCRC-1041; ATCC) and HFFC (300715; Cytion) were purchased, while the third cell line (MoNa) was kindly provided by Dr. Vladimir Rotrekl (Masaryk University), and was originally obtained from the National Tissue Centre (Czech Republic).

Techniques: In Vitro