Human prostate cancer cell lines CWR22Rv1, DU145 and LNCaP (ATCC, Manassas, VA) were cultured in RPMI 1640 (Mediatech, Herndon, VA, #15-041-CV) containing 10% fetal bovine serum (FBS; Quality Biological, Gaithersburg, MD, #100106) and penicillin/streptomycin (Mediatech, Inc., 50 IU/ml and 50 μg/ml, respectively, #30-002-CL). LNCaP cells were cultured in the presence of 0.5 nM dihydrotestosterone (DHT; Sigma, St. Louis, MO, #512-18-6). CWR22Rv1 cells were obtained in 2005 from Dr. Thomas Pretlow (Casewestern Reserve Univ.) and LNCaP and DU145 cells in 2009 from ATCC. AZD1480 (37 (link)) was provided by AstraZeneca. IL-6 producing lentivirus (LV-II-human-IL-6 virus (3.8 × 108 IU/ml) was purchased from Capital Biosciences (Rockville, MD) and recombinant IL-6 was purchased from ProSpec Protein Specialists (Brunswick, NJ, #CYT-213).
Cwr22rv1
Manufactured by American Type Culture Collection
Sourced in United States
The CWR22Rv1 is a human prostate cancer cell line maintained by the American Type Culture Collection (ATCC). It is a widely used model for prostate cancer research. The core function of this cell line is to provide a standardized and renewable source of prostate cancer cells for experimental studies.
Automatically generated - may contain errors
Lab products found in correlation
Lncap, by American Type Culture Collection (51 mentions)
Du145, by American Type Culture Collection (26 mentions)
Rwpe 1, by American Type Culture Collection (9 mentions)
Hek293t, by American Type Culture Collection (9 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (8 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (6 mentions)
Rwpe 1, by Thermo Fisher Scientific (4 mentions)
Rpmi 1640, by Thermo Fisher Scientific (4 mentions)
Mg132, by Merck Group (3 mentions)
L glutamine, by Thermo Fisher Scientific (3 mentions)
Nci h660, by American Type Culture Collection (3 mentions)
Rpmi 1640 media, by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
β actin, by Cell Signaling Technology (2 mentions)
Dihydrotestosterone dht, by Merck Group (2 mentions)
Enzalutamide, by Selleck Chemicals (2 mentions)
R1881, by Merck Group (2 mentions)
Penicillin streptomycin, by Corning (2 mentions)
Cycloheximide (chx), by Merck Group (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
75 protocols using cwr22rv1
1
Prostate Cancer Cell Line Cultivation
2
Characterization of KDM4A and USP1 Interaction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HEK293T cells and PC cell lines CWR22RV1 (hereafter named RV1), LNCAP, VCAP, LN95, C4‐2B, DU145, and PC3 were purchased from ATCC. All cell lines were tested and authenticated by karyotyping analysis on 2 June 2018. Expression plasmids containing pCMV‐Flag‐KDM4A, pCMV‐Flag‐USP1, and pCMV‐Myc‐USP1 were constructed as previously described,18 and pCMV‐Myc‐ USP1 C90S mutant (a deubiquitinating enzyme activity‐disrupted mutant, in which a Cys to Ser point mutation was introduced) was generated using a QuikChange Site‐Directed Mutagenesis Kit (Stratagene) and validated by DNA sequencing. The HA‐ub plasmid and mutation constructs were kindly provided by Professor Jianfei Qi from the University of Maryland Cancer Center.
KDM4A (C37E5) rabbit mAb #5328, USP1 (D37B4) rabbit mAb #8033, c‐Myc (E5Q6W) rabbit mAb #18583, AR (D6F11) XP rabbit mAb #5153, di‐methyl‐histone H3 (Lys9) (D85B4) XP rabbit mAb #4658, and NKX3.1 (D2Y1A) XP rabbit mAb #83700 were purchased from Cell Signaling Technology. Anti‐ub (sc‐8017) Abs were purchased from Santa Cruz Biotechnology. Antibodies against HA (H9658), FLAG (F1804), and β‐actin (A1978) were purchased from Sigma. The USP1 inhibitor ML323 was from Selleck Chemicals. Cycloheximide was purchased from Apexbio.
KDM4A (C37E5) rabbit mAb #5328, USP1 (D37B4) rabbit mAb #8033, c‐Myc (E5Q6W) rabbit mAb #18583, AR (D6F11) XP rabbit mAb #5153, di‐methyl‐histone H3 (Lys9) (D85B4) XP rabbit mAb #4658, and NKX3.1 (D2Y1A) XP rabbit mAb #83700 were purchased from Cell Signaling Technology. Anti‐ub (sc‐8017) Abs were purchased from Santa Cruz Biotechnology. Antibodies against HA (H9658), FLAG (F1804), and β‐actin (A1978) were purchased from Sigma. The USP1 inhibitor ML323 was from Selleck Chemicals. Cycloheximide was purchased from Apexbio.
3
Lentiviral shRNA Screening of LNCaP Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
LNCaP (ATCC CRL-1740) and CWR22Rv1 (ATCC CRL 2505) cells were cultured in RPMI1640 media supplemented with 10% FBS and incubated at 37°C in a humidified incubator containing 5% CO2. DU145 (ATCC HTB-81) and HEK293T (ATCC CRL-3216) cells were cultured in DMEM media supplemented with 10% FBS. LNCaP were infected with aliquots of the DECODE (OpenBiosystems) pooled pGIPZ lentivirus library encoding human genomic shRNAs (13,650 genes targeted in 7 pools of 9,750 shRNA clones/pool) at a multiplicity-of-infection of 1 (RPCI shRNA Core, Irwin Gelman, Director), and then selected for puromycin (2 :g/ml) resistance. Puromycin-resistant cells infected with empty pGIPZ alone served as a negative control.
4
Prostate Cancer Cell Line Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Prostate cancer cell lines, including CWR22Rv1(ATCC CRL-250), PC3 (ATCC CRL-1435), LNCaP (ATCC CRL-1740), DU145 (ATCC HTB-81), C4-2B (ATCC CRL-3315), and immortalized prostate cell line, RWPE-1 (ATCC CRL-11609), were grown in RPMI-1640 supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. For arginine-deprivation condition, RPMI-1640 medium without L-Arginine, L-Leucine, and L-Lysine (US Biological, R8999-03A) supplemented with 10% dialysis fetal bovine serum, 1% penicillin/streptomycin, L-Leucine (50 mg/ml, Sigma), and L-Lysine (40 mg/ml, Sigma) was used. For arginine-stimulation experiment, L-Arginine (200 mg/ml) was freshly added after overnight arginine starvation. Cells were routinely tested for mycoplasma contamination using a PCR-based detection analysis (e-Myco #25233).
5
Cell Culture and Synchronization
6
Comparative analysis of AR-positive prostate cancer cell lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
LNCaP, CWR22Rv1 and VCaP cells were all purchased from ATCC and authenticated prior to conducting the study (see Supplementary Figure S3B for example of authentication). R1-D567 is a TALEN-engineered cell line derivative of the AD-1 cell line that expresses only the AR-V567es receptor isoform1. All cells were maintained in RPMI 1640 media (Sigma) supplemented with 10% foetal calf serum (FCS) and 5% l -glutamine at 37°C and subject to regular mycoplasma testing. Enzalutamide (Selleckchem) and dihydrotestosterone (DHT) (Sigma) were utilised at 10 μM and 10 nM, respectively. PARP inhibitors rucaparib, olaparib and talazoparib were all purchased from Selleckchem and used at 0.5 and 1 μM for assay-specific durations.
7
Cell Culture Conditions for Prostate Cancer Research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We used two different PCa cell lines (C4-2 and CWR22Rv-1; ATCC Cat# CRL-3315, RRID:CVCL_4782 and Cat# CRL-2505, RRID:CVCL_1045), one normal bladder epithelial cell line (SV-HUC; ATCC Cat# CRL-9520, RRID:CVCL_3798), and the 293T cell line. C4-2 and CWR22Rv-1 cells were maintained in RPMI 1640, SV-HUC cells in F-12K media, and 293T cells in DMEM media, all with penicillin (25 units/ml), streptomycin (25 g/ml), 1% L-glutamine, and 10% fetal bovine serum (FBS). For the castration resistant condition, the 10% FBS medium was replaced with phenol red free media containing 10% charcoal-depleted (CD) FBS and 1 nM DHT (this concentration simulates ADT, because it replicates the DHT concentration remaining in the tumors of PCa patients following castration).
8
Cell Culture Conditions for Prostate Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
LNCaP, CWR-22RV1, and PC-3 cells were purchased from ATCC and maintained between passage 9 and 35, in RPMI-1640 (Corning) medium supplemented with 10% fetal bovine serum (FBS) (Cytiva, Marlborough, MA, USA) and penicillin–streptomycin (Corning). R49F cells were kindly gifted to us by Dr Anima Zoubedi and maintained in the presence of 10 μM enzalutamide (Cayman Chemical) in RPMI medium, similar to other prostrate cancer cells. HEK-293 cells were maintained in in DMEM (Corning) medium supplemented with 10% FBS and penicillin–streptomycin.
To maintain basal levels of AR/AR-V7 and TM4SF3, cells were grown in RPMI-1640 or DMEM medium (HEK-293T) without phenol red andl -glutamine, supplemented with 2% dextran charcoal extracted FBS (2% DCC) (Gibco), also referred as deprivation medium, for 48 h before MDM2 inhibitor (MDM2i) treatment or transfections. For androgen treatments, cells were grown in deprivation medium for 48 h, before treatment with ethanol or 10 nM R1881 (Sigma-Aldrich).
To maintain basal levels of AR/AR-V7 and TM4SF3, cells were grown in RPMI-1640 or DMEM medium (HEK-293T) without phenol red and
9
Establishing Prostate Cancer Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Prostate cancer cell lines, CWR-R1, CWR22Rv1 and LNCaP were procured from ATCC (Manassas, VA, USA) and cultured as recommended in RPMI-1640 media containing 10% heat-inactivated fetal bovine serum (FBS, GIBCO) and 1% penicillin-streptomycin (10,000 U/mL, Life Technologies, Carlsbad, CA, USA) at 37 °C and 5% CO2. Chemical synthesis of Galeterone (Gal) and VNPP433-3β was performed as described earlier and dissolved in cell culture-grade DMSO [12 (link)]. The primary antibodies targeting human AR, Mnk1, Mnk2, GAPDH, β-actin, α-tubulin, Histone H3, Ubiquitin and secondary HRP-conjugated anti-rabbit and anti-mouse used in the study were procured from Cell Signaling Technology, USA. Dihydrotestosterone (DHT), MG132 and all fine chemicals were purchased from Sigma Aldrich, St. Louis, MI, USA.
10
Prostate Cancer Cell Line Cultivation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human prostate cancer LNCaP, VCaP,
and CWR22Rv1cells were obtained from the ATCC (Rockville, MD, U.S.A.).
α-T, γ-T and δ-T were from Sigma Co. (St. Louis,
MO, U.S.A.). A mixture of tocopherol γ-TmT was purchased from
the Cogns Corp. (Cincinnati, OH, U.S.A.). The extracellular matrix
Matrigel was obtained from Corning Co. (Tewksbury, MA, U.S.A.). The
various human prostate cancer cells were maintained in RPMI-1640 culture
medium as described in our earlier study.17 (link)
and CWR22Rv1cells were obtained from the ATCC (Rockville, MD, U.S.A.).
α-T, γ-T and δ-T were from Sigma Co. (St. Louis,
MO, U.S.A.). A mixture of tocopherol γ-TmT was purchased from
the Cogns Corp. (Cincinnati, OH, U.S.A.). The extracellular matrix
Matrigel was obtained from Corning Co. (Tewksbury, MA, U.S.A.). The
various human prostate cancer cells were maintained in RPMI-1640 culture
medium as described in our earlier study.17 (link)
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!