colocalization module  (Oxford Instruments)

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

Journal: bioRxiv

Article Title: Mitochondrial carbonic anhydrase-VB inhibition rescues brain endothelial stress and memory in Alzheimer’s disease models

doi: 10.64898/2026.03.16.711716

Figure Lengend Snippet: (A) Staining of cCASP3 and CD31 colocalization in the hippocampus of WT, 3xTG, and 3xTG mice treated with 4ITP (as in Methods). CD31 is represented in red and cCASP3 is represented in green. The %Area of colocalized cCAS3 and CD31 was quantified with HALO imaging software. Original magnification 60x . Quantification on the right. N=4-7 mice/group, mixed male and female. Data represents 5-10 images per mouse and area. Statistical significance was evaluated by One-way ANOVA followed by Tukey post-hoc test. *P< 0.05, **P<0.01. (B) 20x images of WT, 3xTG, and 3xTG + 4ITP cerebral vessels stained with CD31 in red, DAPI in blue. (C ) Quantification of % area of CD31; (D) median blood vessel diameter of WT, untreated 3xTG mice and 3xTG mice treated with 4ITP. (C-D) N=5-6 mice/group, mixed male and female. Data represents 5-10 images per mouse and area. Statistical significance was evaluated by One-way ANOVA followed by Tukey post-hoc test. *P< 0.05, **P<0.01 (E-F) Western blot analysis of TJP occludin and extravasation proteins VCAM-1, ICAM-1 in the (E) hippocampus and (F) cortex of WT, 3xTG, and 3xTG +4ITP. (E, F ) Data represents N=6 mice per group, 2 technical replicates/mouse. Statistical significance was evaluated by Two-way ANOVA followed by Tukey’s post-hoc test (main column effect). *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. Male and female mice are represented by blue and pink dots, respectively.

Article Snippet: All image analysis was conducted using the object colocalization FL module in HALO Image Analysis Platform version 4.2.6399 (Indica Labs, Inc).

Techniques: Staining, Imaging, Software, Western Blot