Journal: Heliyon

Article Title: Upregulation of EphA4 deteriorate brain damage by shifting microglia M1-polarization via NF-κB signaling after focal cerebral ischemia in rats

doi: 10.1016/j.heliyon.2023.e18429

Figure Lengend Snippet: EphA4 Overexpression Aggravated Brain Damage and Neuronal Apoptosis after Ischemia. (A1-A5) Representative MRI images showing infarcted areas after MCAO. (A6) Infarct volume evaluated at day 3 after MCAO. (B1–B5) TUNEL (+) staining (red) plus DAPI (blue) to label apoptotic cells in the boundary zone of ischemic area. Scale Bar = 50 μm (B6) Counting of TUNEL-positive cells in peri-ischemic regions of MCAO rats on postoperative day 3. The results were represented as the mean ± SD, and the differences between groups were compared with ANOVA followed by LSD's post hoc test (n = 5, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns: no significance). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Flow Cytometry method was applied to measure the apoptotic cells with application of Annexin V-Fluorescein Isothiocyanate (FITC) Early Apoptosis Detection Kit (Cell Signaling Technology, USA) following instruction provided.

Techniques: Over Expression, TUNEL Assay, Staining

Journal: Heliyon

Article Title: Upregulation of EphA4 deteriorate brain damage by shifting microglia M1-polarization via NF-κB signaling after focal cerebral ischemia in rats

doi: 10.1016/j.heliyon.2023.e18429

Figure Lengend Snippet: EphA4 Overexpression Increased Apoptosis and Microglia Proliferation Induced by OGD. (A1-A6) Representative data of apoptosis detected by flow cytometry. (B1–B6) Microglia proliferation was tested via EdU (red) staining and DAPI staining (blue). Scale bar = 100 μm. (C–D) Summary of the percentage of apoptotic cells in each group. (E) Counting of EdU-positive microglia cells in co-cultures. The results were represented as the mean ± SD, and the differences between groups were compared with ANOVA followed by LSD's post hoc test (n = 5, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns: no significance). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Flow Cytometry method was applied to measure the apoptotic cells with application of Annexin V-Fluorescein Isothiocyanate (FITC) Early Apoptosis Detection Kit (Cell Signaling Technology, USA) following instruction provided.

Techniques: Over Expression, Flow Cytometry, Staining

Journal: International Journal of Nanomedicine

Article Title: Nanofabrication of methylglyoxal with chitosan biopolymer: a potential tool for enhancement of its anticancer effect

doi: 10.2147/IJN.S78284

Figure Lengend Snippet: Apoptotic effect of Nano-MG and MG on HBL-100 cell line. Notes: ( A ) Flow cytometric analysis of HBL-100 cell line. MG and Nano-MG were incubated for 48 hours at different concentrations. ( B ) Confocal microscopy images (100×) of nuclei stained by Hoechst 33342 dye. Abbreviations: A, area; FITC, fluorescein isothiocyanate; LL, lower left; LR, lower right; MG, methylglyoxal; Nano-MG, MG-conjugated chitosan nanoparticles; UL, upper left; UR, upper right.

Article Snippet: Cells were trypsinized and doubly stained using an annexin V–fluorescein isothiocyanate (FITC) and PI kit (Cell Signaling Technology kit no 6592S) according to the manufacturer’s protocol for flow cytometric analysis of apoptotic populations.

Techniques: Incubation, Confocal Microscopy, Staining

Journal: Environmental Health Perspectives

Article Title: Characterizing the Low-Dose Effects of Methylmercury on the Early Stages of Embryo Development Using Cultured Human Embryonic Stem Cells

doi: 10.1289/EHP7349

Figure Lengend Snippet: Measures of apoptosis in hESCs exposed to MeHg for 24 h and 6 d. (A) Representative image of positive staining of cleaved caspase-3 (CC3) (green), nuclei (DAPI, blue) and α -tubulin (red) and quantification of CC3-positive cells (as a percentage of nuclei) in hESCs exposed to 0 and 200 nM MeHg for 24 h. Scale bar: 100 μ m ; scale bar in the Zoom-in of (A): 10 μ m . (B) Flow cytometry analysis of cell population (percentage) positively stained for Annexin V-FITC and propidium iodide (PI) as measures of early apoptosis and necrosis after 24 h of MeHg treatment, respectively. (C) Expression of CC3 protein after 6 d of MeHg exposure. Each batch was first normalized to its sodium carbonate vehicle control ( 0 nM ) before comparison. Thus, there is no error bar in the 0 nM MeHg group. (D) Flow cytometry analysis of cell population (percentage) positively stained for Annexin V-FITC and PI as measures of early apoptosis and necrosis after 6 d of MeHg treatment, respectively. Data are presented as means ± SEMs with N = 3 for immunostaining and flow cytometry (24 h), N = 4 for western blot, and N = 5 for flow cytometry (6 d). The small circle symbols represent individual values of each experiment. One-way ANOVA was used at each time point to compare between the different doses, and the dose response of the treatment effects was significant for (A) CC3 expression at 24 h and (D) Annexin V-positive cells at 6 d. Dunnett’s post hoc test was then used to compare the treatments to the vehicle control. Significance (*) was set at p < 0.05 . The corresponding numeric data are provided in Table S3. Note: ANOVA, analysis of variance; DAPI, 4′,6-diamidino-2-phenylindole; FITC, fluorescein-5-isothiocyanate; hESC, human embryonic stem cell; MeHg, methylmercury; SEM, standard error of the mean.

Article Snippet: The Annexin V–fluorescein-5-isothiocyanate (FITC) Early Apoptosis Detection Kit (Catalog no. 6592S; Cell Signaling Technology) was used according to the manufacturer’s instructions to detect the effects of MeHg on early apoptosis.

Techniques: Staining, Flow Cytometry, Expressing, Immunostaining, Western Blot

Journal: Environmental Health Perspectives

Article Title: Characterizing the Low-Dose Effects of Methylmercury on the Early Stages of Embryo Development Using Cultured Human Embryonic Stem Cells

doi: 10.1289/EHP7349

Figure Lengend Snippet: Analysis of the cell cycle of hESCs after 6 d of exposure to MeHg. (A) Flow cytometry analysis of cell cycle phases, with a histogram showing the peak for FITC-staining of DNA content as identifications of cell phases and bar graph showing the percentage of cells in different cell cycle phases of hESCs after 6 d of exposure to 0, 10, 50, and 200 nM MeHg. (B) Protein expression of cyclin D1 was measured in hESCs after 6 d of exposure to 0, 5, 10, 50, 100, and 200 nM MeHg. Data are presented as means ± SEMs with N = 3 for each group. For western blots, each batch was first normalized to its sodium carbonate vehicle control ( 0 nM ) before comparison. Thus, there is no error bar in the 0 nM MeHg group. The small circle symbols represent individual values of each experiment. One-way ANOVA was used at each time point to compare between the different doses, and the dose response of the treatment effects was significant. Dunnett’s post hoc test was then used to compare the treatments to the vehicle control. Significance (*) was set at p < 0.05 . The corresponding numeric data are provided in Table S6. Note: ANOVA, analysis of variance; FITC, fluorescein-5-isothiocyanate; hESC, human embryonic stem cell; MeHg, methylmercury; SEM, standard error of the mean.

Article Snippet: The Annexin V–fluorescein-5-isothiocyanate (FITC) Early Apoptosis Detection Kit (Catalog no. 6592S; Cell Signaling Technology) was used according to the manufacturer’s instructions to detect the effects of MeHg on early apoptosis.

Techniques: Flow Cytometry, Staining, Expressing, Western Blot