Journal: International Journal of Molecular Sciences

Article Title: Pilot Proteomic Analysis of Urinary Extracellular Vesicles Supports the “Toxic Urine Hypothesis” as a Vicious Cycle in Refractory IC/BPS Pathogenesis

doi: 10.3390/ijms27010130

Figure Lengend Snippet: Urinary extracellular vesicles from IC/BPS patients enhance NF-κB activity in HEK293T reporter cells and human bladder epithelial cells. ( A ) Schematic representation of the dual-luciferase reporter assay workflow. HEK293T cells were seeded at 0 h, transfected with NF-κB reporter (Firefly luciferase) and internal control (Renilla luciferase) plasmids at 16 h, stimulated with uEVs (50 µg/mL) or TNFα (10 ng/mL) at 32 h, and assessed for luciferase activity at 56 h. pGL3-Basic vector lacking NF-κB binding sites served as a negative control. ( B ) NF-κB activity represented as normalized Firefly/Renilla luciferase ratios. Data are presented as the mean ± SEM. Statistical significance was determined between the indicated groups: *, p < 0.05. ( C ) Here, 50 µg/mL of uEVs from a healthy donor (H) or an IC patient (Pt) was used to treat human bladder epithelial cells (HBlEpC) for 48 h. The protein expressions of 14-3-3ζ, p-NF-κB p65 at serine536, and total NF-κB p65 were determined via Western blot. GAPDH was used as an internal protein loading control. The numbers below the blots indicate expression levels relative to H uEV–treated cells after normalization to GAPDH.

Article Snippet: For the detection of cellular proteins, human bladder epithelial cells (HBlEpC; Cell Applications, Inc., San Diego, CA, USA) were treated with uEVs for 48 h and lysed with RIPA buffer, followed by SDS-PAGE and immunoblot analysis using the indicated antibodies.

Techniques: Activity Assay, Luciferase, Reporter Assay, Transfection, Control, Plasmid Preparation, Binding Assay, Negative Control, Western Blot, Expressing

Journal: Cell reports

Article Title: Cancer Cells Resist Mechanical Destruction in Circulation via RhoA/Actomyosin-Dependent Mechano-Adaptation

doi: 10.1016/j.celrep.2020.02.080

Figure Lengend Snippet: (A) Effects of 2–10 FSS pulses on the viability of freshly isolated prostate cancer cells from tumor-bearing mice (Pten−/− and Pten−/−;Trp53−/−) and on wild-type prostate epithelial cells (****p < 0.0001, two-way ANOVA).

Article Snippet: Primary Bladder Epithelial Cells , ATCC , PCS-420-010.

Techniques: Isolation

Journal: Cell reports

Article Title: Cancer Cells Resist Mechanical Destruction in Circulation via RhoA/Actomyosin-Dependent Mechano-Adaptation

doi: 10.1016/j.celrep.2020.02.080

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Primary Bladder Epithelial Cells , ATCC , PCS-420-010.

Techniques: Virus, Recombinant, shRNA

Journal: British Journal of Cancer

Article Title: H-Ras oncogene counteracts the growth-inhibitory effect of genistein in T24 bladder carcinoma cells

doi: 10.1038/sj.bjc.6602272

Figure Lengend Snippet: The susceptibilities of bladder TCC cell lines and BDEC to growth inhibition by genistein. The growth inhibition curves for BDEC and eight human bladder TCC cell lines are illustrated. To prevent confusion caused by drawing nine curves with error bars in a small plot, we divided the studied cell lines into three groups (panels A – C ) according to their relative growth rates in the presence of increasing genistein concentrations. Panel A : insensitive cell lines include T24 (▿) and primary bladder epithelial cells (□). Panel B : 5637 (▿), BFTC905 (□), HT1197 (○), and J82 (▵) were cell lines that were moderately inhibited. Panel C : relative sensitive cell lines include SCaBER (▵), TCCSUP (▿), and TSGH-8301 (□). These experiments were performed twice with duplicate samples.

Article Snippet: Primary bladder epithelial cells (BDEC) were obtained from BioWhittaker (San Diego, CA, USA) and maintained as recommended by the supplier.

Techniques: Inhibition