Journal: Cell reports

Article Title: Spatial and single-cell transcriptomics reveal the reorganization of cerebellar microglia with aging

doi: 10.1016/j.celrep.2025.116624

Figure Lengend Snippet: (A) Schematic overview of the cellular layers of mouse cerebellum and hippocampus. (B) MERFISH imaging of cerebellum and hippocampus from 3- and 24-month-old mice, colored by major cell types. (C and D) Bar plot showing the changes in microglial proximity to cerebellar (C) and hippocampal (D) cell types in aged versus young mice. Proximity ratios (24 months old/3 months old) for cell-microglia contacts were computed relative to permutation-based null distributions (1,000 iterations), with standard errors estimated by the delta method. Two-tailed z-tests were performed against the null expectation, and p values were corrected using the Benjamini-Hochberg false discovery rate (FDR) method. Bars represent the mean ± SEM. * p < 0.05 and * p < 0.05. (E) MERFISH imaging highlighting microglia (maroon), granule cells distant from microglia (green), granule cells near microglia (blue), and other cells (gray) in the cerebellum and hippocampus of 3- and 24-month-old mice. (F) Stacked bar plot showing the proportion of microglia within the molecular layer, granule layer, and white matter area of the cerebellum in 3- and 24-month-old mice. (G) Representative images of cerebellum from 3-month-old and 24-month-old mice, stained with GABRA6 to label granule cells (green), IBA1 to label microglia (red), and DAPI to label nuclei (blue). Scale bar: 100 μm. (H) Heatmap showing the top differentially expressed genes in microglia from the granule layer (green) compared to the molecular layer (blue) and white matter area (orange) in the aged cerebellum (black).

Article Snippet: GABRA6 , Synaptic Systems , Cat#224603; RRID:AB_2619945.

Techniques: Imaging, Two Tailed Test, Staining

Journal: International Journal of Molecular Sciences

Article Title: Ivy Leaf Dry Extract EA 575 ® Has an Inhibitory Effect on the Signalling Cascade of Adenosine Receptor A 2B

doi: 10.3390/ijms241512373

Figure Lengend Snippet: Influence of EA 575 ® ( A , B ) or the antagonists PSB-603 and SCH 442416 ( C ) on the CRE activation in transiently transfected HEK cells mediated by stimulation with adenosine ( A , C ) or BAY 60-6583 ( B ). Pre-incubation with 40–240 µg/mL EA 575 ® was conducted for 16 h ( A , B ) or with 0.1–10 µM of the antagonists for 2 h ( C ) before cells were stimulated by adding 100 µM adenosine ( A , C ) or 10 µM BAY 60-6583 ( B ). The non-specifically mediated CRE activation was significantly inhibited by pre-incubation with 80–240 µg/mL EA 575 ® ( A ) or 1 µM PSB-603 ( C ), compared to stimulated control cells not pre-incubated with EA 575 ® (SC). SCH 442416 also significantly reduced the CRE activation at a concentration of 10 µM, whereas this effect could not be observed at lower concentrations of both antagonists. Instead, 0.1 µM SCH 442416 slightly increased the CRE activation ( C ). The inhibition of the specific A 2B AR-mediated CRE activation was achieved by pre-incubation with 80–240 µg/mL EA 575 ® ( B ). Influence of the A 2A AR agonist CGS 21680 and, in comparison, BAY 60-6583 and adenosine on the CRE activation in transiently transfected HEK cells ( D ). Stimulation was performed with 1–10 µM CGS 21680, 10 µM BAY 60-6583, or 100 µM adenosine. The A 2A AR-mediated CRE activation was significantly increased by 10 µM CGS 21680 compared with completely untreated control cells (UTC), but to a lesser extent than that mediated by A 2B AR or non-specifically using adenosine ( D ). Data are shown as AUC of the fold change (FC) after stimulation normalised to stimulated control cells not pre-incubated with EA 575 ® (SC) ( A – C ) or completely untreated control cells (UTC) ( D ). Results represent the mean and SEM ( n = 3 independent experiments performed in triplicate, * p < 0.05).

Article Snippet: PSB-603 and CGS 21680 were obtained from Biomol (Hamburg, Germany).

Techniques: Activation Assay, Transfection, Incubation, Concentration Assay, Inhibition

Journal: International Journal of Molecular Sciences

Article Title: Ivy Leaf Dry Extract EA 575 ® Has an Inhibitory Effect on the Signalling Cascade of Adenosine Receptor A 2B

doi: 10.3390/ijms241512373

Figure Lengend Snippet: Influence of EA 575 ® ( A , B ) or the antagonists PSB-603 and SCH 442416 ( C ) on the IL-6 release of Calu-3 cells mediated by stimulation with adenosine ( A , C ) or BAY 60-6583 ( B ). Pre-incubation with 40–240 µg/mL EA 575 ® was conducted for 16 h ( A , B ) or with 0.1–10 µM of the antagonists for 1 h ( C ) before cells were stimulated by adding 100 µM adenosine ( A , C ) or 10 µM BAY 60-6583 ( B ) for another 24 h. The non-specifically mediated IL-6 release was significantly and dose-dependently inhibited by pre-incubation with 80–240 µg/mL EA 575 ® (A) or 1 µM PSB-603 ( C ), compared to stimulated control cells not pre-incubated with EA 575 ® (SC). SCH 442416, however, increased the concentration of IL-6 even further. ( C ). The inhibition of the specific A 2B AR-mediated IL-6 release was achieved by pre-incubation with 40–240 µg/mL EA 575 ® ( B ). Influence of the A 2A AR agonist CGS 21680 and, in comparison, BAY 60-6583 on the IL-6 release of Calu-3 cells ( D ). Stimulation was performed with 1–10 µM CGS 21680 or 10 µM BAY 60-6583 for 24 h. The A 2A AR-mediated IL-6 release was slightly, but neither significantly nor dose-dependently, increased compared to completely untreated control cells (UTC) ( D ). Results represent the mean normalised to stimulated control cells not pre-incubated with EA 575 ® (SC) ( A – C ) or completely untreated control cells (UTC) ( D ) and SEM ( n = 3 independent experiments performed in triplicate, * p < 0.05).

Article Snippet: PSB-603 and CGS 21680 were obtained from Biomol (Hamburg, Germany).

Techniques: Incubation, Concentration Assay, Inhibition