mammary epithelial cell line mcf 10a  (Procell Inc)

Journal: Cancer Reports

Article Title: Regulation of ER Stress‐Associated Gene Expression by Lifeguard in Human Breast Cancer Cells

doi: 10.1002/cnr2.70687

Figure Lengend Snippet: Molecular changes associated with LFG‐specific siRNA treatment in MDA‐MB‐231 and MCF10A cells. (A) LFG mRNA expression measured by qRT‐PCR after 24 and 48 h. Expression was normalized to ACTB and calculated using the 2 −ΔΔCt method with MDA‐MB‐231 control cells as the calibrator. Data represent mean ± SD from three independent experiments. * p < 0.05 versus the corresponding control (Student's t ‐test). (B) Representative fluorescence images showing intracellular Ca 2+ ‐associated fluorescence in MDA‐MB‐231 cells following LFG‐specific siRNA transfection. Images shown are representative of three independent biological experiments ( n = 3). Cells were imaged using identical microscope settings, exposure time, gain, and illumination intensity for all experimental groups. Green fluorescence represents intracellular calcium‐associated fluorescence. Scale bar = 200 μm. (C) Relative expression of selected ER stress‐ and apoptosis‐related genes in MDA‐MB‐231 and MCF10A cells 48 h after LFG‐specific siRNA transfection, determined using the RT 2 Profiler PCR Array. Gene expression was normalized to ACTB (β‐actin) and calculated using the 2 −ΔΔCt method with the corresponding non‐targeting siRNA‐transfected cells as the calibrator. Genes with a fold change ≥ 2 or ≤ −2 were considered differentially expressed. Data are presented as mean ± SD from three independent biological experiments ( n = 3). Statistical comparisons were performed using an unpaired two‐tailed Student's t ‐test, with p < 0.05 considered statistically significant. Error bars represent standard deviation (SD). The asterisk indicates statistical significance compared with the control group ( p < 0.05)

Article Snippet: The non‐tumorigenic human mammary epithelial cell line MCF10A was obtained from ATCC and cultured in DMEM/F12 medium (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 5% horse serum (Sigma‐Aldrich, St. Louis, MO, USA), 100 ng/mL cholera toxin, 20 ng/mL epidermal growth factor, 0.01 mg/mL insulin, and 500 ng/mL hydrocortisone.

Techniques: Expressing, Quantitative RT-PCR, Control, Fluorescence, Transfection, Microscopy, Gene Expression, Two Tailed Test, Standard Deviation