Journal: Nutrients

Article Title: Essential Oils, Pituranthos chloranthus and Teucrium ramosissimum , Chemosensitize Resistant Human Uterine Sarcoma MES-SA/Dx5 Cells to Doxorubicin by Inducing Apoptosis and Targeting P-Glycoprotein

doi: 10.3390/nu13051719

Figure Lengend Snippet: Effects of Pituranthos chloranthus (PC) and Teucrium ramosissimum Desf. (TR) extracts on normal primary human uterine fibroblast cells (HUF) and primary murine Bone Marrow-Derived Macrophages (BMDM) viability. After treatment of primary HUF and murine BMDM with increasing concentrations (0–100 µg/mL) of PC and TR for 72 h, the percentage of viable cells was assessed using the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. ( A ) Dose–response curves of PC-treated HUF (left panel) and TR-treated HUF (right panel). ( B ) Dose–response curves of PC-treated BMDM (left panel) and TR-treated BMDM (right panel). Data are expressed as a mean percentage of control growth ± Standard Deviation (SD) of two representative experiments ( n = 6 replicates per concentration).

Article Snippet: Primary human uterine fibroblast normal cells (HUF) were obtained from the ATCC.

Techniques: Derivative Assay, MTT Assay, Control, Standard Deviation, Concentration Assay

Journal: Nucleic acids research

Article Title: Sequestration of ribosomal subunits as inactive 80S by targeting eIF6 limits mitotic exit and cancer progression.

doi: 10.1093/nar/gkae1272

Figure Lengend Snippet: Figure 9. Ov ere xpression of eIF6 observ ed primarily in high-grade in v asiv e bladder and breast cancers. ( A and B ) Western blot probed for eIF6 in human bladder cancer cell lines and healthy (normal) bladder epithelial cells. β-Tubulin used as loading control. Blots shown in panel (A) were quantitated and eIF6 le v els w ere normaliz ed to loading control and plotted (panel B) as FC o v er eIF6 le v els in normal bladder epithelial cells. Values indicate standard error of the mean from four independent experiments and significant differences for T24 ( P = 0.0065), UMUC3 ( P = 0.0132) and HT1197 ( P = 0.0017) determined by an unpaired t wo-t ailed t -test. ( C ) Images represent eIF6 expression in patient-derived benign and high-grade tumors by immunohistochemistry using anti-eIF6 antibody. Enlarged inset shows the presence of eIF6 in nucleoli and cytoplasm in high-grade cancers. ( D ) Images shown in panel (C) were quantitated and eIF6 expression in patient-matched benign tissues relative to high-grade cancers (six patients) were plotted. Significant differences were determined using an unpaired t wo-t ailed t -test ( P = 0.0063). ( E ) Plot shows eIF6 expression in unmatched benign tissues, high-grade and low-grade bladder cancers derived from patients. ( F and G ) Western blot represents high levels of eIF6 in high-grade invasive human triple negative breast cancer cell line. β-Tubulin used as loading control. Blots shown in panel (F) were quantitated and eIF6 levels were normalized to loading control and plotted (G) as FC o v er eIF6 le v els in normal (healthy) HME-1 cells. Values indicate standard error of the mean from four independent experiments and significant differences for MDA-MB-231 ( P = 0.001) determined by an unpaired t wo-t ailed t -test.

Article Snippet: The human bladder cancer cell lines HT1376, HT1197, UM-UC-3 and TCCSUP were maintained in EMEM media with 10% FBS and Penn / Strep, T-24 cell lines were maintained in McCoy’s 5A medium with 10% FBS and Penn / Strep and primary bladder epithelial (A / T / N): normal human (BdEC) cells were maintained in bladder epithelial cell basal medium (ATCC) supplemented with bladder epithelial g c i g b D W C [ a p 1 fl d t ( m a p c w m ( b l o β S 0 p B S C C e d s i S B m B s a ( S B a p a m t w s i i H 7 w f v D ow nloaded from https://academ ic.oup.com /nar/advance-article/doi/10.1093/nar/gkae1272/7933602 by guest on 30 D ecem ber 2024 rowth factors.

Techniques: Western Blot, Control, Expressing, Derivative Assay, Immunohistochemistry

Journal: International Journal of Molecular Sciences

Article Title: High-Content Imaging and Machine Learning Classify Phenotypical Change in Coronary Artery Endothelial Cells Caused by BPS

doi: 10.3390/ijms27073259

Figure Lengend Snippet: Representative high-content microscopy images of human coronary artery endothelial cells (HCAEC) exposed to vehicle control (CTRL) or 0.1 µM Bisphenol S (BPS) for 96 h and stained using the PhenoVue Cell Painting assay. For each condition, a representative field acquired at 40× magnification and a higher-magnification inset are shown. Rows correspond to the individual fluorescence channels: Hoechst 33342 (nuclei), PhenoVue Fluor 488 Concanavalin A (endoplasmic reticulum and intracellular membranes), PhenoVue 512 nucleic acid stain (RNA/nucleoli), PhenoVue Fluor 555 wheat germ agglutinin (plasma membrane), PhenoVue 641 mitochondrial stain (mitochondria), and the merged image. White boxes represent the part of the image used for the related inset. Scale bar: 50 µm, 40× objective.

Article Snippet: Primary human coronary artery endothelial cells (HCAEC; ATCC ® PCS-100-020TM, Innovation, VA, USA) were cultured according to the supplier’s recommendations.

Techniques: Microscopy, Control, Staining, Fluorescence, Clinical Proteomics, Membrane

Journal: Cell reports

Article Title: SILAC Analysis Reveals Increased Secretion of Hemostasis-Related Factors by Senescent Cells

doi: 10.1016/j.celrep.2019.08.049

Figure Lengend Snippet: (A) A schematic outlining the strategy for SILAC-based identification of SASP proteins. See for time lines of how cells were made quiescent or senescent and how quiescent and senescent conditioned media (CM) were produced. (B) Reactome pathway analysis of proteins secreted at >2-fold levels by senescent compared with quiescent HCA2 cells. Pathways related to hemostasis are identified by boxes. (C) Heatmap showing individual levels of proteins, identified by SILACs, that participate in hemostasis and are secreted by quiescent (QUI) and senescent (SEN [IR]) HCA2 normal human fibroblasts. (D) CM were collected from quiescent (QUI) and senescent (SEN [IR]) cells, concentrated, and analyzed using western blotting for the indicated SASP factors. Cells remaining on the plate at the time of CM collection were counted for normalization. Mindin protein levels and Ponceau S staining were used as loading controls. Fold changes are mean ± SEM. *p < 0.05 and **p < 0.01. See also and .

Article Snippet: WI-38 and IMR-90 are normal primary human embryonic lung fibroblasts (obtained from the American Type Culture Collection).

Techniques: Multiplex sample analysis, Produced, Western Blot, Staining

Journal: Cell reports

Article Title: SILAC Analysis Reveals Increased Secretion of Hemostasis-Related Factors by Senescent Cells

doi: 10.1016/j.celrep.2019.08.049

Figure Lengend Snippet: (A) mRNA was extracted from HCA2 cells made quiescent (QUI) or induced to senesce by doxorubicin (SEN [DOXO]), and analyzed using qPCR for the indicated genes, using actin mRNA for normalization. The data are shown in a.u., and the bars show mean ± SEM. *p < 0.05 and **p < 0.01, two-tailed, unpaired t test. (B) CM were collected from cells treated as in (A), concentrated and analyzed using western blotting for the indicated SASP factors. Cells were counted for normalization. Mindin protein levels and Ponceau S staining were used as loading controls. (C) IMR-90 fibroblasts were depleted of mtDNA by culture in ethidium bromide until senescent (21 days) (labeled as MiDAS). RNA was extracted and analyzed using qPCR for the indicated factors. Heatmap represents individual values for three experiments. *p < 0.05, one-way ANOVA. See also .

Article Snippet: WI-38 and IMR-90 are normal primary human embryonic lung fibroblasts (obtained from the American Type Culture Collection).

Techniques: Two Tailed Test, Western Blot, Staining, Labeling

Journal: Cell reports

Article Title: SILAC Analysis Reveals Increased Secretion of Hemostasis-Related Factors by Senescent Cells

doi: 10.1016/j.celrep.2019.08.049

Figure Lengend Snippet:

Article Snippet: WI-38 and IMR-90 are normal primary human embryonic lung fibroblasts (obtained from the American Type Culture Collection).

Techniques: Recombinant, Modification, Sequencing, Multiplex sample analysis, Labeling, Enzyme-linked Immunosorbent Assay, Sandwich ELISA, Reverse Transcription, Software

Journal: Cancers

Article Title: Preclinical Evaluation of the Novel Small-Molecule MSI-N1014 for Treating Drug-Resistant Colon Cancer via the LGR5/β-catenin/miR-142-3p Network and Reducing Cancer-Associated Fibroblast Transformation

doi: 10.3390/cancers12061590

Figure Lengend Snippet: Cancer-associated fibroblasts (CAFs) increased the oncogenic properties of colon cancer cells. ( A ) Insert illustrates the co-culture system of DLD1 and HCT116 colorectal cancer (CRC) cells with Normal fibroblasts (NF) and CAFs. Cell viability assay showed increased 5-fluorouracil (5-FU) resistance in CAF-educated DLD1 and HCT116 cells, compared to their naïve counterparts. Flow cytometric analysis of DLD1 and HCT116 cells co-cultured with CAFs. Increased CD44 + cell population ( B ) and side population ( C ) in both DLD1 and HCT116 cells, 48 h post CAF culture. Enhanced colony-forming ( D ), migratory ( E ), and tumor sphere-generating ( F ) abilities in both CRC cell lines post CAF co-culture. ** p < 0.01, *** p < 0.001.

Article Snippet: The DLD1 and HCT116 human colon cancer cell lines and normal fibroblasts (NFs, ATCC® PCS-201-018) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA) and were cultured according to the vendor’s recommended conditions.

Techniques: Co-Culture Assay, Viability Assay, Cell Culture