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

Journal: Frontiers in Pharmacology

Article Title: Extracellular vesicle-mediated preconditioning of osteoblasts by ER+ breast cancer cells: implications for the bone metastatic niche

doi: 10.3389/fphar.2026.1844422

Figure Lengend Snippet: T47D cells exhibit enhanced extracellular vesicle production. (A) Schematic illustrating extracellular vesicle (EV) biogenesis pathways, including exosome release through the endosomal-multivesicular body pathway and microvesicle (MV) shedding via outward budding from the plasma membrane. Schematic adapted from Nashed, M. (2026) on BioRender. (B) Representative histogram from nanoparticle tracking analysis (NTA) showing EV production rate and size distribution of EVs for T47D breast cancer cells and MCF10A cells as a control (n = 3 per group). Particle concentration is normalized to the max peak for each experimental group. Shaded area shows the standard error of the mean (SEM) for all histograms. (C) Maximum production rate in the exosome regime (30–150 nm) for T47D and MCF10A cells. (D) Maximum production rate in the MV regime (shown here as 150–500 nm) for T47D and MCF10A cells. All data and error bars in the bar graphs represent mean ± SEM from n = 3 independent experiments. Statistical significance was determined using an unpaired two-tailed Student’s t-test to calculate represented p -values; n.s indicates not significant.

Article Snippet: Human MCF10A breast epithelial cells (ATCC, CRL-10317) were cultured in DMEM/F12 medium (ATCC, 30–2006) supplemented with 5% horse serum (ATCC, 30–2030), human recombinant epidermal growth factor (EGF; Gibco, PHG0311) at a final concentration of 20 ng/mL, hydrocortisone (Sigma-Aldrich, H0888) at 0.5 μg/mL, cholera toxin (Sigma-Aldrich, C8052) at 100 ng/mL, and human recombinant insulin (Gibco, 12-585-014) at 10 μg/mL.

Techniques: Clinical Proteomics, Membrane, Analysis, Control, Concentration Assay, Two Tailed Test