Journal: The FASEB Journal

Article Title: Mechanical Phenotyping of MG63s Following Vibrational Stimulation

doi: 10.1096/fj.202504885R

Figure Lengend Snippet: Mechanical stimulation setup and high‐throughput whole cell mechanical analysis. (A) T75 flasks are attached to the nanovibration device by adhesive magnets. The device utilizes piezo actuators to vibrate the plate vertically. The nanovibration device is calibrated to output sinusoidal motion of 1000‐Hz frequency and 30‐nm amplitude. (B) Bioreactor's calibration profile using laser interferometry presenting an output signal of 1000 Hz frequency and 30.42 ± 1.75 nm (Mean ± SD) amplitude. (C) Cell's mechanical properties are tested utilizing Real‐Time Deformability Cytometry. The samples are tested under zero stress (reservoir) and under stress (channel). High‐throughput technique allows testing n = 5000 under 2 min assuming 1 × 10 6 cells/mL.

Article Snippet: Osteosarcoma immortalized cells, MG63, were cultured in T75 cell culture flasks (Fisher Scientific, 10 364 131) using Dulbecco's Modified Eagle Medium (DMEM) (VWR, 392–0415) supplemented with 10% (v/v) fetal bovine serum (FBS) (Fisher Scientific, 17 593 595), non‐essential amino acids (Thermo Fisher, 11 140 050), and Penicillin–Streptomycin (Pen/Strep) (Merck, P4458) in a humidified incubator at 37°C in 5% CO 2 .

Techniques: High Throughput Screening Assay, Adhesive, Cytometry

Journal: Journal of Cellular and Molecular Medicine

Article Title: EN2 Regulates Pancreatic Cancer Initiation, Progression, and Epithelial‐Mesenchymal Transition Through the Notch Signalling Pathway

doi: 10.1111/jcmm.71158

Figure Lengend Snippet: The expression of EN2 in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. (A), Protein expression of EN2 in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. Crude proteins were isolated, and EN2 expression was measured by Western blot analysis. β‐Actin was used as a loading control. (B), Expression of EN2 mRNA in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. RNA was isolated, and EN2 expression was measured by q‐RT‐PCR. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. *, # and % = significantly different from HPNE ( p < 0.05). (C), Expression of EN2. Immunocytochemistry was performed to examine EN2 expression in HPNE, PANC‐1, and AsPC‐1 cells.

Article Snippet: Human pancreatic cancer stem cells (CSCs) were obtained from Celprogen and cultured in a well‐defined medium according to the manufacturer's instructions.

Techniques: Expressing, Isolation, Western Blot, Control, Reverse Transcription Polymerase Chain Reaction, Immunocytochemistry