The cellular variants and their culture or transduction procedures are thoroughly described in previous research [6 (link),7 (link)]. In brief, bladder cancer RT-112, HT-1376 and CAL-29 cell lines were purchased from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ, Brunswick, Germany), incubated in 37 °C under 5% CO2 and cultured in either RPMI-1640 (RT-112) or DMEM (HT-1376, CAL-29) medium supplemented with fetal bovine serum, Antibiotic-Antimycotic and L-glutamine. Lentiviral transduction was performed via different overexpression systems for WWOX and AP-2α/γ in polybrene-containing starvation medium. Antibiotic clone selection using puromycin (WWOX) or G418 (AP-2α/γ) was preceded by exchange of viral medium to full medium. The following stable transductants were obtained for each cell line: Control/control (KK); Control/AP-2α↑ (KA); Control/AP-2γ↑ (KC); WWOX↑/control (WK); WWOX↑/AP-2α↑ (WA); WWOX↑/AP-2γ↑ (WC). In order to identify genes whose expression is inversely regulated by WWOX or AP-2 and alleviated during their combination, in this research the WA and WC variants were considered as a rescue experiment to KK: these are models where the level of WWOX/AP-2 expression is balanced to some extent compared to KA, KC or WK. Established comparisons together with their purpose are summarized in Figure 1 .
Rt112
Manufactured by Leibniz Institute DSMZ
Sourced in Germany, United States
The RT112 is a laboratory instrument designed for real-time PCR (polymerase chain reaction) analysis. It is used for the amplification and quantification of nucleic acid sequences. The RT112 provides accurate and reproducible results for various applications, including gene expression analysis, pathogen detection, and DNA/RNA quantification.
Automatically generated - may contain errors
Lab products found in correlation
Vm cub1, by Leibniz Institute DSMZ (6 mentions)
Tccsup, by American Type Culture Collection (4 mentions)
Ht1197, by American Type Culture Collection (4 mentions)
Ht1376, by American Type Culture Collection (4 mentions)
Sw780, by American Type Culture Collection (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Scaber, by American Type Culture Collection (2 mentions)
Scaber, by Leibniz Institute DSMZ (2 mentions)
Kmbc2, by Japanese Collection of Research Bioresources Cell Bank (2 mentions)
Umuc3, by American Type Culture Collection (2 mentions)
Bc 3c, by Leibniz Institute DSMZ (2 mentions)
Ku 19 19, by Leibniz Institute DSMZ (2 mentions)
Cal 29, by Leibniz Institute DSMZ (2 mentions)
Bftc 905, by Leibniz Institute DSMZ (2 mentions)
Sw 1710, by Leibniz Institute DSMZ (2 mentions)
Mfm 223, by Leibniz Institute DSMZ (2 mentions)
Minimum essential medium (mem), by Thermo Fisher Scientific (1 mentions)
Nih3t3, by American Type Culture Collection (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
17 protocols using rt112
1
Bladder Cancer Cell Line Manipulation
2
Bladder Cancer Cell Line Cultivation
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The bladder cancer cell lines TCCSUP (HTB-5), 5637 (HTB-9), RT4 (HTB-2), T24 (HTB-4), ScaBER (HTB-3), HT1197 (CRL-1473), HT1376 (CRL-1472), SW780 (CRL-2169), NIH/3T3 (CRL-1658), and SK-BR-3 (HTB-30) were obtained from ATCC (Manassas, VA, USA). RT112 was obtained from DSMZ, Germany. Metastatic lines of T24 - T24T, FL3, and SLT3 were gifted by Dr. Michael Nickerson (Division of Cancer Epidemiology & Genetics, NCI). The bladder cancer cell lines 253 J, UMUC-5, and UMUC-1 were a kind gift from Dr. David McConkey (Johns Hopkins University). MGH-U3 was obtained from Massachusetts General Hospital. RT4v6 was a gift from Dr. Peter Black (University of British Columbia). All cells were grown in minimum essential media (MEM) (Life technologies) supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin and 1% GlutaMAX (Life technologies). All cells were incubated in a humidified incubator at 37 °C with 5% CO2 for growth.
3
Characterization of Bladder Cancer Cell Lines
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The human bladder cancer-derived cell lines RT112, UM-UC-5, UM-UC-9, VM-CUB-1, 5637, 647 V and HT1376 were obtained from DSMZ (Heidelberg, Germany). MGH-U3 cells were kindly provided by Dr. Francisco X. Real (CNIO, Madrid). MGH-U3, UM-UC-5, UM-UC-9, VM-CUB-1, 647 V and HT1376 cells were cultured in DMEM, whereas RT112, and 5637, cells were cultured in RPMI. Media were supplemented with 10% foetal bovine serum (FBS) (ThermoFisher Scientific, Courtaboeuf, France). Cells were kept at 37 °C, under an atmosphere containing 5% CO2. The identity of the cell lines used was checked by analysing genomic alterations on comparative genomic hybridization arrays (CGH array) and sequencing genes known to be mutated: RAS, TP53, FGFR3 and PIK3CA. The cells were routinely checked for mycoplasma contamination.
4
Bladder Cancer Cell Line Cultivation
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The human bladder cancer-derived cell lines UMUC7, UMUC14, RT112, RT4, VMCUB1, SCaBER, UMUC6, T24 and HT1197 were obtained from DSMZ (Heidelberg, Germany). KMBC2 cells were purchased from JCRB cell bank (Japan). KMBC2 cells were cultured at 37 °C in an atmosphere of 5% CO2 in Ham F12 medium, RT112 and RT4 cells were cultured in RPMI medium and all the other cells were cultured in DMEM medium. All cell media were supplemented with 10% fetal bovine serum (FBS). Cells were routinely tested for mycoplasma contamination.
5
Bladder Cancer Cell Line Characterization
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Bladder cancer cell lines HT1376 (RRID CVCL_1292), T24 (CVCL_0554), and 5637 (CVCL_0126) were obtained from ATCC, and RT112 (CVCL_1670) and VM-CUB1 (CVCL_1786) were obtained from DSMZ. Cell lines were authenticated by the vendors and tested for mycoplasma prior to use. These were grown in Dulbecco’s 90% MEM (4.5 g/L glucose) with penicillin/streptomycin (100 U/mL) and 10% fetal bovine serum (MilliporeSigma) in 5% CO2 incubators. GSK-126 (HY-13470), alpelisib (HY-15244), and dactolisib (HY-50673) were obtained from MedChem Express. Pictilisib (S1065), MG132 (S2619), MAK-683, EPZ-6438, and CPI-1205 were obtained from Selleckchem. Captisol (RC-0C7-020) was obtained from CyDex Pharmaceuticals.
6
Establishing Chemotherapy-Resistant Bladder Cancer Cell Lines
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HT‐1197, HT‐1376, J82, RT4, SW 780, TCCSUP, and UM‐UC‐3 were purchased from ATCC (Manassas, VA, USA). 647‐V, BC‐3C, BFTC‐905, CAL‐29, KU‐19‐19, RT‐112, SW‐1710 and VM‐CUB1 were purchased from DSMZ (Braunschweig, Germany). EJ138, U‐BLC1, UM‐UC‐9 and UM‐UC‐14 were purchased from ECACC (Salisbury, UK). KMBC‐2 and T24 were purchased from JCRB Cell Bank (Osaka, Japan). BOY‐12E, and JMSU‐1 were provided by the RIKEN BRC (Tsukuba, Japan). These cell lines were cultured according to the guidelines from the suppliers.
To generate chemotherapy‐resistant cell lines, UM‐UC‐14 and RT‐112 cell lines were exposed to adriamycin and gemcitabine, respectively, whose concentrations were gradually increased up to 100 and 1000 ng/mL, respectively. Adriamycin‐resistant UM‐UC‐14 and gemcitabine‐resistant RT‐112 cell lines were maintained in the culture medium containing 50 ng/mL adriamycin and 1000 ng/mL gemcitabine, respectively.
To generate chemotherapy‐resistant cell lines, UM‐UC‐14 and RT‐112 cell lines were exposed to adriamycin and gemcitabine, respectively, whose concentrations were gradually increased up to 100 and 1000 ng/mL, respectively. Adriamycin‐resistant UM‐UC‐14 and gemcitabine‐resistant RT‐112 cell lines were maintained in the culture medium containing 50 ng/mL adriamycin and 1000 ng/mL gemcitabine, respectively.
7
Bladder Cancer Cell Lines and Normal Urothelial Cells
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The human bladder cancer-derived cell lines RT112, 5637, KK47, and SCaBER were obtained from DSMZ (Heidelberg, Germany). MGH-U3, KK47 and SD48 cell lines were provided by Yves Fradet (CRC, Quebec), Jennifer Southgate laboratory (previously of Cancer Research Unit, St James’s University Hospital, Leeds, UK), and Henri Mondor Hospital (Créteil, France), respectively. The L1207 cell line was derived fromt tumour T1207 [67 (link)]. RT112 and 5637 cells were cultured in RPMI medium, L1207 were cultured in DMEM-F12 and all the other cell lines were cultured in DMEM medium. All cell media were supplemented with 10% fetal bovine serum (FBS). We used Normal human urothelium (NHU) cells obtained from normal ureter urothelium from healthy kidney donors from Foch hospital and were cultured as previously described [68 ]. NHU derived from ureter have been shown to be transcriptionally very similar to NHU derived from bladder [69 (link), 70 (link)]. All cells were cultured at 37 °C in an atmosphere of 5% CO2 and were routinely tested for mycoplasma contamination.
8
Characterization of Bladder Cell Lines
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The uroepithelial cell line SV-HUC-1 (RRID: CVCL_3798) and bladder cancer cell lines UM-UC-3 (RRID: CVCL_1783), SW780 (RRID: CVCL_1728), RT4 (RRID: CVCL_0036), and 5637 (RRID: CVCL_0126) were purchased from the Cell Bank of Chinese Academy of Sciences (Shanghai, China), RT112 (RRID: CVCL_1670) was purchased from DSMZ, T24T (RRID: CVCL_M892) and EJ (RRID: CVCL_2893) were gifted by Dr. Guosong Jiang of Wuhan Union Hospital. All cell lines were cultured in appropriate complete medium and maintained at 37 °C in 5% CO2 condition.
All cell lines have been authenticated using short tandem repeat profiling within the last 3 years and tested for mycoplasma contamination by DAPI staining. All experiments were performed with mycoplasma-free cells.
All cell lines have been authenticated using short tandem repeat profiling within the last 3 years and tested for mycoplasma contamination by DAPI staining. All experiments were performed with mycoplasma-free cells.
9
Characterization of Bladder Cancer Cell Lines
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Bladder cancer cell lines 5637, HT1197, HT1376, J82, RT4, SCaBER, SW780, T24, TCCSUP and UM-UC3 were purchased from ATCC (Manassas, VA, USA). 647V, BC-3C, BFTC-905, CAL-29, JMSU-1, KU-19-19, RT-112, SW1710 and VM-CUB1 were purchased from DSMZ (Riverdale, MD, USA). 253J, SV-HUC, TSU-PR1, UM-UC14 and WH were obtained from Dr. Jer-Tsong Hsieh (University of Texas Southwestern, Dallas, TX, USA). Bladder cell lines 94-10, 96-1, 97-1, 97-7, 97-18, 97-24, DSH1, JO’N, SD, VM-CUB2 and VM-CUB3 were obtained from Dr. Margaret A. Knowles (University of Leeds, Leeds, UK). Cell lines were re-authenticated via short tandem repeat (STR) analysis using the Cell-ID-system (G9500, Promega, Nacka, Sweden), and products analyzed using an Applied Biosystems 3130 Genetic Analyzer (San Francisco, CA, USA). Cell lines were cultured according to the manufacturer recommendations. We performed tests for Mycoplasma sp. and cell identification by Single Nucleotide Polymorphism (SNP) using the MycoSEQ detection kit (Applied Biosystems, Waltham, MA, USA).
10
Urothelial Cancer Cell Line Characterization
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The human urothelial cancer cell lines RT4 (p53 wild type), HT-1197 (p53 wild type), TCC-SUP (mutant p53), T-24 (mutant p53), were purchased from ATCC (Manassas, VA, USA) [19 (link)]. RT112 (p53 wild type) cell line was purchased from DSMZ (Braunschweig, Germany) [19 (link)]. 486p cell line (unknown p53 mutational status) was previously generated and characterized by Elliott et al. from grade IV TCC of the bladder metastatic to a supraclavicular node of 61-year-old white male patient [20 (link), 21 (link)]; ethics and consent statements can be found in the corresponding references [20 (link), 21 (link)]. 486p cells were kindly provided by the Urology department of University Medical Center Hamburg-Eppendorf, Hamburg, Germany. Culture conditions are described in the Additional file 1 .
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