Journal: Science Advances

Article Title: Glycerol-mediated nose-to-brain codelivery of anti–IL-17 and anti-CD73 antibodies enhances immunotherapy for melanoma brain metastases

doi: 10.1126/sciadv.adx7966

Figure Lengend Snippet: ( A and B ) Mice underwent brain fluorescence imaging at 0, 0.5, 1, 2, 4, 8, 12, 24, and 48 hours postintranasal delivery of Cy7-labeled a–IL-17 (Cy7–a–IL-17) and Cy5-labeled a-CD73 (Cy5–a-CD73). ( C and D ) Quantitative analysis of the fluorescence intensity for Cy7–a–IL-17 (C) and Cy5-a-CD73 (D) ( n = 3). ( E and F ) Quantitative analysis of the fluorescence intensity for Cy7–a–IL-17 (E) and Cy5–a-CD73 (F) in isolated brains at 2 hours ( n = 3). ( G ) Representative fluorescence microscopy images of brain sections 2 hours after intranasal delivery or intravenous injection of FITC–a–IL-17 (green) or Cy5–a-CD73 (red). Nuclei were counterstained with DAPI (blue). Scale bars, 20 μm. ( H to K ) Quantitative analysis of fluorescence intensity in the olfactory bulb (H), cortex (I), hippocampus (J), and cerebellum (K) ( n = 3). ( L ) Representative images of brain immunofluorescence were captured 2 hours after intranasal delivery or intravenous injection of FITC-IgG (green) to observe the distribution of antibodies in brain tissue sections. Blood vessels were stained with CD31 (red). Scale bars, 20 μm. All statistics are expressed as means ± SD. Statistical significance was assessed using one-way analysis of variance (ANOVA) with Fisher’s LSD test, where * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. ns, nonsignificant.

Article Snippet: For precise cellular localization, the sections were subjected to immunofluorescence staining: The vascular endothelium was labeled with an anti-CD31 primary antibody (catalog no. GB12063-100, Servicebio, Hubei, China), visualized with a Cy3-conjugated secondary antibody, and counterstained with DAPI for nuclei.

Techniques: Fluorescence, Imaging, Labeling, Isolation, Microscopy, Injection, Olfactory, Immunofluorescence, Staining

Journal: The Journal of General Physiology

Article Title: Beat-locked ATP microdomains in the sinoatrial node map a Ca 2+ -timed energetic hierarchy and regional pacemaker roles

doi: 10.1085/jgp.202513874

Figure Lengend Snippet: Superior SA node myocytes exhibit elevated diastolic ATP and metabolic flux compared with the inferior region. (A) 3D segmented maximum-intensity projection of a whole-mount SA node immunolabeled for CD31 (vasculature, red) and cyto-iATP (myocytes, green). The dashed line denotes the boundary between superior and inferior regions. (B) Image-processing workflow illustrating merged maximum-intensity projections, binary segmentation masks, and extraction of grayscale cyto-iATP signals used for quantitative analysis. (C) Mean cyto-iATP fluorescence intensity per myocyte, grouped by region ( N = 5 mice per region), reporting expression levels of the EGFP-tagged cyto-iATP sensor. (D) Live confocal imaging of cyto-iATP signals showing representative line-scan images and corresponding normalized fluorescence traces (F/F 0 ) from superior and inferior regions. (E and F) Summary quantification of cyto-iATP signal mass rate (E) and estimated diastolic [ATP] i (F). P values are shown above comparisons. Large circles denote per-animal means; small circles indicate individual biological replicates. N represents the number of independent mice.

Article Snippet: For immunolabeling, SA nodes were incubated for 48 h at 4°C with a goat anti-mouse CD31 primary antibody (1:50, AF3628; R&D Systems).

Techniques: Immunolabeling, Extraction, Fluorescence, Expressing, Imaging