Journal: G3: Genes|Genomes|Genetics

Article Title: Maternal Proteins That Are Phosphoregulated upon Egg Activation Include Crucial Factors for Oogenesis, Egg Activation and Embryogenesis in Drosophila melanogaster

doi: 10.1534/g3.118.200578

Figure Lengend Snippet: Maternal knockdown of 15 genes led to developmental arrest in stage 1 and/or early stage 2 of embryogenesis. 2-4 hr old embryos from knockdown females, stained for tubulin (green) and DNA (red). For each set of images, the one on the right shows a close-up view of the spindle (marked by the arrow on the left). Embryos from AttP2 control females mated with ORP2 males are used as positive controls. Scale bars: embryo 50 µm, nucleus 5 µm.

Article Snippet: Mouse anti-γtubulin (Boster Biological Technologies, Pleasanton, CA, catalog #MA1114) was reconstituted in 1ml of PBS and used at a dilution of 1:100.

Techniques: Knockdown, Staining, Control

Journal: G3: Genes|Genomes|Genetics

Article Title: Maternal Proteins That Are Phosphoregulated upon Egg Activation Include Crucial Factors for Oogenesis, Egg Activation and Embryogenesis in Drosophila melanogaster

doi: 10.1534/g3.118.200578

Figure Lengend Snippet: Maternal knockdown of 15 new maternal effect factors led to centrosome abnormalities. (A) Representative images of metaphase spindles in early embryos with normal (AttP2) or abnormal (maternal RNAi knockdowns) centrosome arrangement. 2-4hr old embryos are stained for αtubulin (green), and γtubulin (blue). Scale bar: 5µm. Gamma tubulin signals are marked by white arrows. (B) Proportion of maternal knockdown or control embryos with normal, supernumerary, acentrosomal spindles or spindle with single centrosome. A summary of the centrosome activity in the embryos with knockdown of these 15 genes can be found in Table S7.

Article Snippet: Mouse anti-γtubulin (Boster Biological Technologies, Pleasanton, CA, catalog #MA1114) was reconstituted in 1ml of PBS and used at a dilution of 1:100.

Techniques: Knockdown, Staining, Control, Activity Assay

Journal: G3: Genes|Genomes|Genetics

Article Title: Maternal Proteins That Are Phosphoregulated upon Egg Activation Include Crucial Factors for Oogenesis, Egg Activation and Embryogenesis in Drosophila melanogaster

doi: 10.1534/g3.118.200578

Figure Lengend Snippet: Polar bodies with abnormal morphology were observed in knockdown embryos for 11 genes. Significant portions of 0.5-1 hr embryos with maternal knockdown of Dlig-I (83% abnormal, n = 23), mcm3 (56% abnormal, n = 36), spc105r (96%, n = 23), plu (100%, n = 30), spindly (53% abnormal, n = 32), aub (57%, n = 28), mri (62% abnormal, n = 37), 14-3-3ε (93% abnormal, n = 29), mod(mdg4) (58% abnormal, n = 24), ball (88%, n = 24) and PyK (13%, n = 31) had polar bodies with abnormal morphology. Tubulin is shown in green. DNA is shown in red. Scale bars: 5µm.

Article Snippet: Mouse anti-γtubulin (Boster Biological Technologies, Pleasanton, CA, catalog #MA1114) was reconstituted in 1ml of PBS and used at a dilution of 1:100.

Techniques: Knockdown

Journal: iScience

Article Title: Activating FcγR function depends on endosomal-signaling platforms

doi: 10.1016/j.isci.2023.107055

Figure Lengend Snippet: IRAP deletion affects the endosomal recruitment of active Syk to FcγRI (A and B) Wild-type (WT) or IRAP-deficient (KO) BM-DCs were incubated with anti-mouse FcγRI (clone AT152-9), followed by crosslinking with non-labeled anti-rat IgG at 4°C. After removal of excess antibodies, the cells were shifted for the indicated time points at 37°C, fixed and stained for endogenous FcγRI (green) and γ-chain (red) in (A) and for γ-chain (green) and Syk phosphorylated on Tyrosine 352 (pSyk) (red) in (B). The pictures show representative images from three independent experiments and the graphs show colocalization between FcγRI and the γ-chain (A) and colocalization between the γ-chain and phosphorylated, active Syk (B). Each dot represents a cell. Data are represented as mean ± SEM. Scale bars = 5 μm. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001.

Article Snippet: Anti-γ chain , MBL , Clone 1D6, Cat# M-191-3.

Techniques: Incubation, Labeling, Staining

Journal: International Journal of Molecular Sciences

Article Title: Exploring the Physical and Biological Aspects of BNCT with a Carboranylmethylbenzo[ b ]acridone Compound in U87 Glioblastoma Cells

doi: 10.3390/ijms232314929

Figure Lengend Snippet: DSB quantification using the ɣ-H2AX assay (up); ( A ) represents the nuclei stained by DAPI, ( B ) the foci, pink dots, identified by TXR. The number of MN per BN cell is also presented for CMBA (down) ( C ). The error bars represent the standard deviation of the three independent experiments. The images were obtained using 64× magnification.

Article Snippet: Then, cells were incubated with an anti-γ-H2AX primary antibody (mouse anti-γ-H2AX (ser139), Stressgen, bioreagents Corp., Canada) at 2 μg/mL for 1 h. After being washed twice with 1% BSA in PBS, cells were incubated with a FITC-conjugated anti-mouse secondary antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at 1 mg/mL for 1 h, followed by three washing steps with PBS.

Techniques: Staining, Standard Deviation

Journal: Biology

Article Title: Observation of Histone H2AX Phosphorylation by Radiation-Induced Bystander Response Using Titanium Characteristic X-ray Microbeam

doi: 10.3390/biology12050734

Figure Lengend Snippet: Accumulation of γ-H2AX in Ti K X-ray microbeam-irradiated and non-irradiated HeLa cells. Three targeted cell nuclei were microbeam irradiated at 4 Gy. MBIR (microbeam irradiation) is a fluorescent image captured at the time of microbeam irradiation. Before irradiation, cell nuclei were stained with Hoechst 33258 solution for targeted irradiation of cell nuclei. The red squares are the irradiation positions set by the irradiation software. White dotted circles in the merged images indicate irradiated cell nuclei after fixation and immunofluorescent staining. Bars represent 30 μm.

Article Snippet: WI-38 cells were fixed as described previously [ ] and then incubated overnight at 4 °C with 1% BSA in T-PBS containing anti-γ-H2AX and anti-p53-binding protein 1 (53BP1) antibodies (PC712, Merck) diluted at 1:500.

Techniques: Irradiation, Staining, Software

Journal: Biology

Article Title: Observation of Histone H2AX Phosphorylation by Radiation-Induced Bystander Response Using Titanium Characteristic X-ray Microbeam

doi: 10.3390/biology12050734

Figure Lengend Snippet: Induction of bystander cells after Ti K X-ray microbeam irradiation of the nuclei of three cells in the center of the field of view at a dose of 4 Gy. Unirradiated HeLa cells with γ-H2AX fluorescence intensity higher than the average + σ of control cells were determined to be bystander cells. The error bars represent standard errors of the mean (SEM) based on the four to nine independent experiments. p -values (* p < 0.05, ** p < 0.01, *** p < 0.001) calculated with Tukey’s multiple comparison test.

Article Snippet: WI-38 cells were fixed as described previously [ ] and then incubated overnight at 4 °C with 1% BSA in T-PBS containing anti-γ-H2AX and anti-p53-binding protein 1 (53BP1) antibodies (PC712, Merck) diluted at 1:500.

Techniques: Irradiation, Fluorescence