Journal: Frontiers in Pharmacology

Article Title: Molecular Mechanisms Underlying the Inhibition of Proliferation and Differentiation by Florfenicol in P19 Stem Cells: Transcriptome Analysis

doi: 10.3389/fphar.2022.779664

Figure Lengend Snippet: FLO inhibits the proliferation and pluripotency of P19SCs. (A) P19SC viability in response to FLO-treatment as detected by CCK-8 method. The results are expressed as the average cell viabilities from three independent experiments. (B) Effect of different doses of FLO on LDH release of P19SCs when exposed for 48 h; ns , not significant. (C) Immunoblot analysis of multiple doses of FLO on protein levels of pluripotent markers (Sox2 and Oct4) in P19SCs treated for 48 h.

Article Snippet: The medium was removed, and 100 μL PBS containing 10 μL CCK-8 solution (TransGen Biotech, China) was added into each well and maintained at 37°C.

Techniques: CCK-8 Assay, Western Blot

Journal: Gland Surgery

Article Title: Tumor-derived mesenchymal progenitor cell-related genes in the regulation of breast cancer proliferation

doi: 10.21037/gs-23-387

Figure Lengend Snippet: Comparison of cell proliferation, migration, and secretion abilities of MSCs and MPCs. (A) The cell proliferation was measured via eFluor670 dye at 0, 24, 48, and 72 hours for MSCs and MPCs. (B) MSC and MPC proliferation indices of eFluor670 dye (n=5; *P<0.05, MSC vs. MPC). (C) MSC and MPC proliferation indices of CCK8 assay at 0, 24, 48, and 72 hours (n=5; ***P<0.001, MSC vs. MPC). (D) Cell scratch assay to detect the migration ability of MPCs and MSCs (n=5). (E) Detection of cellular inflammatory factors according to qRT-PCR (n=5; **P<0.01; ***P<0.001). MSC, mesenchymal stem cell; MPC, mesenchymal progenitor cell; IL-6, interleukin 6; CXCL12, C-X-C motif chemokine ligand 12; FGF2, fibroblast growth factor 2; TGF-β, transforming growth factor β; IGF-1, insulin-like growth factor 1; CCK8, Cell Counting Kit 8; qRT-PCR, quantitative real-time polymerase chain reaction.

Article Snippet: MSCs and MPCs were seeded into 96-well plates and processed at 0, 48, and 72 h. CCK8 (10 μL) reagent (ScienCell, Carlsbad, CA, USA) was added to each well, and the cell was cultured again for 2 h; the absorbance was measured at 450 nm with a microplate reader, the experimental results were recorded, and a growth curve was drawn.

Techniques: Comparison, Migration, CCK-8 Assay, Wound Healing Assay, Quantitative RT-PCR, Cell Counting, Real-time Polymerase Chain Reaction

Journal: Gland Surgery

Article Title: Tumor-derived mesenchymal progenitor cell-related genes in the regulation of breast cancer proliferation

doi: 10.21037/gs-23-387

Figure Lengend Snippet: STOM-overexpressing MPCs secrete inflammatory factors while promoting breast cancer proliferation. (A) Plasmid constructs mapping of LAMA5 , CCBE1 , and STOM . (B) qRT-PCR mRNA expression of target genes in MPCs transfected with overexpression plasmids (n=3; *P<0.05; ***P<0.001 target group compared with the control group). (C) qRT-PCR showed upregulation of CXCL12 and FGF2 expression after upregulation of the STOM gene (n=5; ***P<0.001 vs. control). (D) qRT-PCR showed upregulation of IGF expression after upregulation of the CCBE1 gene (n=5; ***P<0.001 vs. control). (E) qRT-PCR showed upregulation of FGF1 expression after upregulation of the LAMA5 gene (n=5; ***P<0.001 vs. control). (F-H) CCK8 assay indicated proliferation of MPCs overexpressing LAMA5 , CCBE1 , and STOM (n=5; ***P<0.001 vs. control). (I-K) CCK8 assay indicated proliferation of breast cancer MDA-MB-231 cells promoted by MPCs overexpressing STOM after coculture, and the overexpression of CCBE1 and LAMA5 did not have this pro-proliferative effect (n=5; ***P<0.001 vs. control). (L) CCK8 assay indicated proliferation of breast cancer MCF-7 cells promoted by MPCs overexpressing STOM after coculture (n=3; ***P<0.001 vs. control). MSC, mesenchymal stem cell; MPC, mesenchymal progenitor cell; qRT-PCR, quantitative real-time polymerase chain reaction; STOM, stomatin; CCBE1, collagen and calcium binding EGF domains 1; LAMA5, laminin subunit alpha 5; CXCL12, C-X-C motif chemokine ligand 12; FGF2, fibroblast growth factor 2; FGF1, fibroblast growth factor 1; IGF, insulin-like growth factor.

Article Snippet: MSCs and MPCs were seeded into 96-well plates and processed at 0, 48, and 72 h. CCK8 (10 μL) reagent (ScienCell, Carlsbad, CA, USA) was added to each well, and the cell was cultured again for 2 h; the absorbance was measured at 450 nm with a microplate reader, the experimental results were recorded, and a growth curve was drawn.

Techniques: Plasmid Preparation, Construct, Quantitative RT-PCR, Expressing, Transfection, Over Expression, Control, CCK-8 Assay, Real-time Polymerase Chain Reaction, Binding Assay