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: Journal of Clinical Laboratory Analysis

Article Title: hsa_circ_0119412 overexpression promotes cervical cancer progression by targeting miR‐217 to upregulate anterior gradient 2

doi: 10.1002/jcla.24236

Figure Lengend Snippet: circ_0119412 downregulation suppressed cervical cancer cell malignant behaviors and tumor growth in animal models. (A) The efficiency of circ_0119412 overexpression was checked by RT‐qPCR. (B) Cell proliferation affected by circ_0119412 overexpression was assessed by CCK‐8 assay. (C) Cell migration affected by circ_0119412 overexpression was assessed by transwell assay. (D) Cell adhesion affected by circ_0119412 overexpression was assessed by MTT assay. (E) The role of circ_0119412 in vivo was assessed by animal study. *p < 0.05, **p < 0.001 relative to oe‐NC

Article Snippet: CCK‐8 reagent (Cwbio) was used to incubate cells in different wells at the indicated time points (24, 48, 72 or 96 h post‐seeding).

Techniques: Over Expression, Quantitative RT-PCR, CCK-8 Assay, Migration, Transwell Assay, MTT Assay, In Vivo

Journal: Journal of Clinical Laboratory Analysis

Article Title: hsa_circ_0119412 overexpression promotes cervical cancer progression by targeting miR‐217 to upregulate anterior gradient 2

doi: 10.1002/jcla.24236

Figure Lengend Snippet: miR‐217 upregulation reversed the effects caused by circ_0119412 overexpression. (A) The expression of miR‐217 in Hela and SiHa cells transfected with oe‐circ, miR‐217 mimic, or oe‐circ+miR‐217 mimic was checked by RT‐qPCR. In these transfected cells, (B‐D) cell proliferation, migration, and adhesion were monitored by CCK‐8, transwell, or MTT assay, respectively, **p < 0.001 relative to oe‐NC; & p < 0.05, && p < 0.001 relative to mimic‐NC; # p < 0.05, ## p < 0.001 relative to oe‐circ+mimic

Article Snippet: CCK‐8 reagent (Cwbio) was used to incubate cells in different wells at the indicated time points (24, 48, 72 or 96 h post‐seeding).

Techniques: Over Expression, Expressing, Transfection, Quantitative RT-PCR, Migration, CCK-8 Assay, MTT Assay

Journal: Journal of Clinical Laboratory Analysis

Article Title: hsa_circ_0119412 overexpression promotes cervical cancer progression by targeting miR‐217 to upregulate anterior gradient 2

doi: 10.1002/jcla.24236

Figure Lengend Snippet: miR‐217 inhibited cervical cancer cell malignant behaviors by degrading AGR2. (A) The expression of AGR2 protein in Hela and SiHa cells transfected with oe‐AGR2, miR‐217 mimic, or oe‐AGR2+miR‐217 mimic was measured by western blot. In these transfected cells, (B‐D) cell proliferation, migration, and adhesion were investigated by CCK‐8, transwell, and MTT assay, respectively, **p < 0.001 relative to oe‐NC; & p < 0.05 and && p < 0.001 relative to mimic‐NC; # p < 0.05, ## p < 0.001 relative to oe‐AGR2+mimic

Article Snippet: CCK‐8 reagent (Cwbio) was used to incubate cells in different wells at the indicated time points (24, 48, 72 or 96 h post‐seeding).

Techniques: Expressing, Transfection, Western Blot, Migration, CCK-8 Assay, MTT Assay

Journal: Annals of Translational Medicine

Article Title: TGR5 overexpression mediated by the inhibition of transcription factor SOX9 protects against hypoxia-/reoxygenation-induced injury in hippocampal neurons by activating Nrf2 / HO-1 signaling

doi: 10.21037/atm-22-5225

Figure Lengend Snippet: The elevation of TGR5 contributes to HT22 cell viability after H/R. (A) RT-qPCR and (B) western blots were used to determine TGR5 expression in the HT22 cells in the absence or presence of H/R treatment. (C) RT-qPCR and (D) western blot analyses of the overexpression efficacy of pcDNA3.1-TGR5 plasmid. The viability of the H/R-treated HT22 cells was assessed by (E) CCK-8 assays. (F) LDH release was measured by a LDH assay kit. *, P<0.05; **, P<0.01; ***, P<0.001. H/R, hypoxia/reoxygenation; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; LDH, lactate dehydrogenase.

Article Snippet: The cells were cultivated for another 2 h at 37 ℃ after the addition of 10 µL of CCK-8 solution (Beijing Transgen Biotech Co., Ltd.).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Over Expression, Plasmid Preparation, CCK-8 Assay, Lactate Dehydrogenase Assay, Real-time Polymerase Chain Reaction

Journal: Annals of Translational Medicine

Article Title: TGR5 overexpression mediated by the inhibition of transcription factor SOX9 protects against hypoxia-/reoxygenation-induced injury in hippocampal neurons by activating Nrf2 / HO-1 signaling

doi: 10.21037/atm-22-5225

Figure Lengend Snippet: The upregulation of SOX9 reverses the inhibitory role of TGR5 in H/R-triggered HT22 cell injury. The viability of H/R-treated HT22 cells was assessed by (A) CCK-8 assays; (B) LDH release was measured by the LDH assay kit. Corresponding kits were used to examine the levels of SOD, GSH-Px, and MDA. The apoptotic ability of the H/R-treated HT22 cells was evaluated by (C) TUNEL assays (magnification, 200×). (D) IF staining was used to detect cyto-c release (magnification, 200×). (E) Western blots were used analyze the protein levels of Bcl-2, Bax, cleaved caspase-3, and cleaved-PARP. *, P<0.05; **, P<0.01; ***, P<0.001. H/R, hypoxia/reoxygenation; NC, negative control; CCK-8, Cell Counting Kit-8; LDH, lactate dehydrogenase; SOD, superoxide dismutase; GSH-Px, glutathione peroxidase; MDA, malondialdehyde; C-Caspase3, cleaved caspase3; C-PARP, cleaved-PARP.

Article Snippet: The cells were cultivated for another 2 h at 37 ℃ after the addition of 10 µL of CCK-8 solution (Beijing Transgen Biotech Co., Ltd.).

Techniques: CCK-8 Assay, Lactate Dehydrogenase Assay, TUNEL Assay, Staining, Western Blot, Negative Control, Cell Counting

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