Journal: Nature communications

Article Title: LEADeR role of miR-205 host gene as long noncoding RNA in prostate basal cell differentiation.

doi: 10.1038/s41467-018-08153-2

Figure Lengend Snippet: Fig. 5 LEADR regulates basal–luminal differentiation. a Bright-field images showing morphological changes occurring in RWPE-1 (left) and PrEC (right) cells (day 3) upon LEADR silencing by siLEADR (top) or gapLEADR (bottom). Scale bar, 50 µm. Full-size images reported in Supplementary Fig. 4c. b qRT-PCR showing changes in basal and luminal cytokeratins in RWPE-1 or PrEC cells upon LEADR silencing by siLEADR (top) or gapLEADR (bottom). Mean + s.d. plotted. c Western blots showing changes in basal/luminal cytokeratins in RWPE-1 (left) or PrEC (right) cells upon LEADR silencing by siLEADR (top) or gapLEADR (bottom). Vinculin used as loading control. d Immunofluorescence showing cytoplasmic re-localization of p63 (green) upon LEADR silencing in RWPE-1 cells by siLEADR (top) or gapLEADR (bottom). Nuclei counterstained with DAPI (blue). Scale bar, 50 µm. Full-size images are reported in Supplementary Fig. 4d. e Western blots showing changes in Androgen Receptor (AR) expression in PrEC cells upon LEADR silencing by siLEADR (left) or gapLEADR (right). GAPDH used as loading control. f Immunofluorescence showing AR (red) expression in PrEC cells upon LEADR silencing by siLEADR, in the presence or absence of simultaneous DHT stimulation. Nuclei counterstained with DAPI (blue). Scale bar, 50 µm. Full-size images reported in Supplementary Fig. 4g. g ELISA-based quantification of PSA in the conditioned media of PrEC cells silenced for LEADR expression by siLEADR (±DHT). Mean + s.d. (n = 2) plotted. h Heatmap (bottom) of the normalized enrichment score (NES) for luminal and basal custom gene sets (obtained from gene- expression data of frankly basal and luminal prostate cells from the indicated datasets) in RWPE-1 cells upon knockdown of LEADR and/or miR-205 (the latter abrogated using an antisense LNA-modified oligomer, LNA205). Bar plot of differential luminal/basal NES is reported (top). Mean + s.d. (n = 4) plotted. i Plot showing propensity of wild type (wt), LEADR-overexpressing and CRISPRed RWPE-1 cells to differentiate toward luminal phenotype (measured by qRT-PCR and expressed as KRT18/KRT5 ratio) upon culturing in media with increasing differentiative potential (n = 3). ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001 (Student’s t test). Source data are provided as a Source Data file, together with n of all experiments

Article Snippet: The normal primary prostate epithelial cells (PrEC) were grown in PrEC basal medium (ATCC; PCS-440-030) supplemented with PrEC growth kit (ATCC; PCS-440-040).

Techniques: Quantitative RT-PCR, Western Blot, Control, Expressing, Enzyme-linked Immunosorbent Assay, Gene Expression, Knockdown

Journal: World Journal of Surgical Oncology

Article Title: Quercetin inhibits prostate cancer by attenuating cell survival and inhibiting anti-apoptotic pathways

doi: 10.1186/s12957-018-1400-z

Figure Lengend Snippet: Quercetin reduces cell viability and induces apoptosis in PCa cells. Normal prostate epithelial cells PrEC and PCa cells (LNCaP, DU-145, PC-3) were treated with quercetin, and MTT assay was used to determine cell viability ( a ). The EC 50 was calculated from the equation of the line of best fit. PCa cells were treated with 40 μM (EC 50 ) of quercetin, and vehicle-treated controls were stained with FITC-conjugated Annexin V and 7-AAD. Data was acquired by FACS and analyzed using FlowJo software ( b ). Dot plot shows percent (%) of early apoptotic (lower right quadrate) and late apoptotic cells (right upper quadrate). Apoptosis induced by quercetin (shown in top bar diagram) as well as induced necrotic cells (bottom bar diagram) were measured by paired t test to show significance compared to controls

Article Snippet: Normal prostate epithelial cells (PrEC), with materials purchased from ATCC, were cultured in basal medium with cell growth kit containing the following: 6 mM L-glutamine, 0.4% Extract P, 1.0 μM epinephrine, 0.5 ng/mL rhTGFα, 100 ng/mL hydrocortisone, 5 μg/mL rh insulin, and 5 μg/mL Apo-transferrin.

Techniques: MTT Assay, Staining, Software

Journal: Genome Biology

Article Title: epiG: statistical inference and profiling of DNA methylation from whole-genome bisulfite sequencing data

doi: 10.1186/s13059-017-1168-4

Figure Lengend Snippet: WGBS and NOMe-seq data sets

Article Snippet: PrEC cells were grown in Prostate Epithelial Cell Basal Medium (ATCC) supplemented with Prostate Epithelial Cell Growth Kit (ATCC).

Techniques: