rt4 human transitional cell papilloma cell line  (ATCC)

Journal: Clinical Epigenetics

Article Title: Repression of the PRELP gene is relieved by histone deacetylase inhibitors through acetylation of histone H2B lysine 5 in bladder cancer

doi: 10.1186/s13148-022-01370-z

Figure Lengend Snippet: Reduced cell viability associated with induced PRELP gene expression in bladder cancer cells. PRELP gene expression was induced by the addition of doxycycline (DOX, 1 μg/ml) in the lentiviral expression system. Expression of myc-tagged PRELP protein was analyzed using whole-cell extracts from RT4 ( A ) and J82 ( B ) cells with or without DOX. The left side indicated the protein size marker. Alpha-tubulin; loading control. Cell viability of RT4 ( C ) and J82 ( D ) cells was evaluated using cell counting kit (CCK)-8 assays. Black lines; without DOX, Red lines; with DOX. Error bars indicate biological replicates ( n = 3). Statistical analysis was performed using paired Student’s t test. * P < 0.05, **** P < 0.0001

Article Snippet: Human urinary bladder transitional cell papilloma cell lines RT4 (HTB-2) and J82 (HTB-1) were purchased from the American Type Culture Collection (Manassas, VA USA).

Techniques: Gene Expression, Expressing, Marker, Control, Cell Counting, CCK-8 Assay

Journal: Clinical Epigenetics

Article Title: Repression of the PRELP gene is relieved by histone deacetylase inhibitors through acetylation of histone H2B lysine 5 in bladder cancer

doi: 10.1186/s13148-022-01370-z

Figure Lengend Snippet: Restoration of PRELP gene expression by HDAC inhibitors in bladder cancer cells. RT-qPCR analyses of PRELP (gray bars) gene expression in RT4 ( A ) and J82 cells ( B ). The cells were treated with either DMSO or increasing concentration of BIX01294 (0, 0.625, 1.25, 2.5, 5, and 10 μM), UNC0638 (0, 0.625, 1.25, 2.5, 5, and 10 μM), EPZ011989 (0, 0.625, 1.25, 2.5, 5, and 10 μM), UNC1999 (0, 0.625, 1.25, 2.5, 5, and 10 μM), 5-azacytidine (0, 0.625, 1.25, 2.5, 5, and 10 μM), TSA (0, 0.25, 0.5, 1.0, and 2 μM), and SAHA (0, 0.3125, 0.625, 1.25, 2.5, and 5 μM) for 72 h. Data are presented as the mean ± standard deviation (SD) of three technical replicates. The x -axis shows the fold increase relative to DMSO-treated control. The y -axis shows the name and concentration of each inhibitor. The red line indicates a fold increase of 1

Article Snippet: Human urinary bladder transitional cell papilloma cell lines RT4 (HTB-2) and J82 (HTB-1) were purchased from the American Type Culture Collection (Manassas, VA USA).

Techniques: Gene Expression, Quantitative RT-PCR, Concentration Assay, Standard Deviation, Control

Journal: Clinical Epigenetics

Article Title: Repression of the PRELP gene is relieved by histone deacetylase inhibitors through acetylation of histone H2B lysine 5 in bladder cancer

doi: 10.1186/s13148-022-01370-z

Figure Lengend Snippet: The HDAC inhibitor, SAHA, restores the expression of the PRELP gene through H2B acetylation at the promoter regions in the bladder cancer cells. A Genomic positions (Region 1 and Region 2) of the two PCR-amplified DNAs fragments are shown. B ChIP-qPCR was performed using various histone antibodies at the PRELP gene promoter region (Region 1) in RT4 cells treated with DMSO or 5 μM SAHA for 24 h. C Same as ( B ), except that region 2 was similarly analyzed. D The acetylation status of H2BK5 in the promoter region (Region 1 and Region 2) of PRELP gene in RT4 cells treated with two inhibitors, C646 (5 μM), SGC-CBP30 (5 μM), in the presence of SAHA (5 μM). Black; DMSO, Red; SAHA, Blue; SAHA with C646, Green; SAHA with SGC-CBP30. E H2BK5ac ChIP-qPCR in J82 cells. The ChIP enrichment (red bars) is normalized against the values obtained with the same antibodies in DMSO-treated control cells (black bars). Bars represent the mean ± SD of three technical replicates or mean ± standard error of the mean (SEM) of three biological replicates. Statistical analysis was performed using paired Student’s t test. * P < 0.05, ** P < 0.01

Article Snippet: Human urinary bladder transitional cell papilloma cell lines RT4 (HTB-2) and J82 (HTB-1) were purchased from the American Type Culture Collection (Manassas, VA USA).

Techniques: Expressing, Amplification, ChIP-qPCR, Control

Journal: Clinical Epigenetics

Article Title: Repression of the PRELP gene is relieved by histone deacetylase inhibitors through acetylation of histone H2B lysine 5 in bladder cancer

doi: 10.1186/s13148-022-01370-z

Figure Lengend Snippet: Combinatory effect of cisplatin and SAHA on viability of bladder cancer cells. Cells were treated with cisplatin alone or in combination with SAHA. Cisplatin and SAHA were used at a concentration range of 0, 0.2, 0.5, 1.0, 2.0, 5.0 μg/ml and 0, 0.5, 1.0, 2.5, 5.0, 10.0 μM, respectively. Cell viability of RT4 ( A ) and J82 ( B ) cells was evaluated using cell counting kit (CCK)-8 assays. We set the viability of cells with no inhibitor as 100% ( n = 4). y -axis shows cell viability (%), and x -axis shows the concentration of cisplatin. Normalized isobologram of RT4 ( C ) and J82 ( D ) show cisplatin and SAHA synergism as described in “ ”

Article Snippet: Human urinary bladder transitional cell papilloma cell lines RT4 (HTB-2) and J82 (HTB-1) were purchased from the American Type Culture Collection (Manassas, VA USA).

Techniques: Concentration Assay, Cell Counting, CCK-8 Assay