Journal: Scientific Reports

Article Title: Newly identified Gon4l/Udu-interacting proteins implicate novel functions

doi: 10.1038/s41598-020-70855-9

Figure Lengend Snippet: Confocal images for the cellular co-localization analysis of Udu and its interacting proteins. For co-localization analysis, Flag-tagged Udu-P/S construct was co-transfected individually with each Myc-tagged construct, including Bap1, Dnmt1, Thoc1, and Cry3a. Anti-FLAG M2 monoclonal antibody and rabbit anti-MYC antibody were used. Alexa Fluor 488-conjugated anti-mouse secondary antibody and Alexa Fluor 564-conjugated anti-rabbit secondary antibody were then applied. Green fluorescence indicates Udu-P/S, while red fluorescence indicates the expression of Myc-tagged proteins. Blue color is DAPI used for nuclear counterstain. Merged images show the nuclear co-localization of Udu and the interacting proteins.

Article Snippet: SlowFad Diamond Antifade Mountant with DAPI (Thermo Fisher Scientific) was used for nuclear counterstain and mounting.

Techniques: Construct, Transfection, Fluorescence, Expressing

Journal: Current biology : CB

Article Title: A conserved PDZ Binding Motif in aPKC interacts with Par-3 and mediates cortical polarity

doi: 10.1016/j.cub.2019.12.055

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Brains were stored in SlowFade Diamond with DAPI (Invitrogen, S36964 ) for at least 24 hours before imaging.

Techniques: Diagnostic Assay, Recombinant, Construct, Variant Assay, Expressing, Blocking Assay, Plasmid Preparation, Clone Assay, Transformation Assay, Software

Journal: International Journal of Molecular Sciences

Article Title: Multiple Irradiation Affects Cellular and Extracellular Components of the Mouse Brain Tissue and Adhesion and Proliferation of Glioblastoma Cells in Experimental System In Vivo

doi: 10.3390/ijms222413350

Figure Lengend Snippet: Co-culture of primary glial cultures from irradiated mouse brains with GBM cells. ( A ) Microphotograph of the co-culture of U87 GBM cells with glial cells from the control or irradiated brain tissues. Magnification ×200, scale bar 500 µm. ( B ) Quantitative analysis of a number of U87 cells per 0.5 × 0.5 cm square (Zeiss LSM710 tile-scan + CellProfiler 4.1.3). * p < 0.05. ( C ) Immunofluorescence analysis of PG core proteins (decorin, brevican) and GAG content (CS, HS) in primary glial cells co-cultured with U87 GBM cells or triple-irradiated or both. Visualization of the studied antigens with Alexa Fluor 488 secondary antibodies (green), U87-RFP cells (red) and DAPI (blue). Magnification ×400,scale bar 100 µm. ( D ). Quantitative analysis of the PG core proteins and GAG content in the glial cultures from control and irradiated mouse brain tissues (CellProfiler4.1.3. software). ANOVA, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: To analyze the adhesion of tumor cells to the surface of organotypic slices, 12,500 cells in 10 μL were applied onto each slice, incubated for 2 h at 37 °C and 5% CO 2 , washed in PBS, and fixed in 10% neutral buffered formalin at 4 °C for 16 h. To assess the proliferation of tumor cells and their invasion into organotypic slices, 5000 cells in 10 μL were applied onto each slice, incubated for 7 days at 37 °C and 5% CO 2 , then washed with PBS and fixed in 10% neutral buffered formalin at 4 °C for 16 h. After fixation, organotypic slices-U87-RFP co-cultures were washed three times in PBS, transferred onto microscope slides and covered with a coverslip using SlowFade Diamond medium with DAPI (Thermo Scientific, Waltham, MA, USA).

Techniques: Co-Culture Assay, Irradiation, Control, Immunofluorescence, Cell Culture, Software

Journal: International Journal of Molecular Sciences

Article Title: Multiple Irradiation Affects Cellular and Extracellular Components of the Mouse Brain Tissue and Adhesion and Proliferation of Glioblastoma Cells in Experimental System In Vivo

doi: 10.3390/ijms222413350

Figure Lengend Snippet: Co-culture of organotypic brain tissue slices ex vivo with GBM cells. ( A ) Effects of multiple irradiation on PGs’ expression in brain tissue. Real-time RT–PCR analysis, intensity of the amplified DNA fragments normalized to that of Gapdh , bars represent the mean ± SD from triplicate experiments, Student t-test (OriginPro 8.5). ( B ) Overall transcriptional activity of PG core protein-coding genes in brain tissue from control and irradiated mouse brains. The stacked columns compare the contribution of each value to a total across categories. ( C ) Confocal microscopy of U87-RFP cells seeded on the control and irradiated brain slices. Cells nuclei are stained with DAPI. Scale bar 500 μm. ( D ) Quantitative analysis of the U87-RFP cells on the control and irradiated brain tissues. ANOVA and post-hoc Fisher’s LSD test, * p < 0.05, ** p < 0.01.

Article Snippet: To analyze the adhesion of tumor cells to the surface of organotypic slices, 12,500 cells in 10 μL were applied onto each slice, incubated for 2 h at 37 °C and 5% CO 2 , washed in PBS, and fixed in 10% neutral buffered formalin at 4 °C for 16 h. To assess the proliferation of tumor cells and their invasion into organotypic slices, 5000 cells in 10 μL were applied onto each slice, incubated for 7 days at 37 °C and 5% CO 2 , then washed with PBS and fixed in 10% neutral buffered formalin at 4 °C for 16 h. After fixation, organotypic slices-U87-RFP co-cultures were washed three times in PBS, transferred onto microscope slides and covered with a coverslip using SlowFade Diamond medium with DAPI (Thermo Scientific, Waltham, MA, USA).

Techniques: Co-Culture Assay, Ex Vivo, Irradiation, Expressing, Quantitative RT-PCR, Amplification, Activity Assay, Control, Confocal Microscopy, Staining