Journal: Scientific Reports

Article Title: Production and control of coagulation proteins for factor X activation in human endothelial cells and fibroblasts

doi: 10.1038/s41598-020-59058-4

Figure Lengend Snippet: Graphs depicting co-localized detection of intracellular coagulation factors with Golgi and ER proteins. Intensities were measured from red and green channels along a single line in merged images of HUVECs stained simultaneously with antibodies against coagulation proteins and antibodies either to Golgi or PDI. The intensities were measured along the white line shown on the merged images (left panels) and the corresponding graphs (right panels). Fluorescence intensity is on the y-axis and distance (in pixels) is on the x-axis. ( a ) Prothrombin (red) plus PDI (green), line = 10 µm ( b ) FVII (red) plus Golgi (green), line = 7 µm ( c ) FIX (red) plus Golgi (green), line = 11 µm and ( d ) FX (red) plus PDI (green), line = 9 µm. HUVECs were imaged at 100× and nuclei were detected with DAPI. Images were selected from 4–16 similar images for each coagulation protein. The specific detection antibodies are listed in the Fluorescence colocalization measurement section of the Methods. The region of interest (ROI) is the line where the intensities were measured.

Article Snippet: The degree of similarity between detection in the red and green channels in merged images was determined using the fluorescence colocalization module within IP Lab software (Scanalytics, Inc.).

Techniques: Coagulation, Staining, Fluorescence

Journal: iScience

Article Title: Dopaminergic neurons are vulnerable to dysregulation of YEATS2-dependent calcium homeostasis

doi: 10.1016/j.isci.2026.115855

Figure Lengend Snippet: Pharmacological attenuation of SOCE restores DAergic synaptic integrity in dYEATS2 -deficient flies (A) Experimental scheme. Flies expressing membrane-tethered mCD8-GFP and dYEATS2 RNAi specifically in dopaminergic neurons ( Ddc>mCD8-GFP>dYEATS2-IR ) were reared on standard medium supplemented with vehicle or the Orai inhibitor BTP2 (YM-58483) at 1 μM or 10 μM. Adult flies were transferred to fresh vials containing the same treatments, and heads were dissected at 5 days post-eclosion for confocal imaging or RNA extraction. (B) Representative confocal images (posterior→anterior orientation) showing functionally active DAergic neurons identified by co-localization of mCD8-GFP (membrane marker expressed under Ddc-GAL4 driver) and tyrosine hydroxylase (TH) immunoreactivity. Central brain boundaries are indicated by dotted lines; major DA clusters (PAL, PAM, PPL1, PPM3) are highlighted with red dashed circles. Scale bars, 250μm. (C) Quantification of EGFP-TH co-localization (number of co-localized puncta) was performed on 10 independent brains per condition using the colocalization module in CellSense (Olympus). Bars show mean ± SEM; BTP2 treatment at both 1 μM and 10 μM significantly increased the number of EGFP-TH co-localizing spots relative to untreated dYEATS2 -IR animals. (D) Transcript levels of selected dYEATS2 -responsive genes ( Gαq , trpL , vMAT , and DD2R ) were measured from dissected adult heads following vehicle or BTP2 treatment to assess whether SOCE inhibition modulates these transcriptional changes. Gene expression was determined by reverse transcription quantitative PCR (RT-qPCR). Data are presented as mean ± SEM. Statistical significance was assessed by one-way ANOVA with Šidák’s and Tukey’s post hoc tests, respectively; p < 0.05 was considered significant.

Article Snippet: Scale bars, 250μm. (C) Quantification of EGFP-TH co-localization (number of co-localized puncta) was performed on 10 independent brains per condition using the colocalization module in CellSense (Olympus).

Techniques: Expressing, Membrane, Imaging, RNA Extraction, Marker, Inhibition, Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative RT-PCR