Journal: bioRxiv

Article Title: Parkinson’s disease risk factors are expressed at brain barriers

doi: 10.1101/2025.10.20.683561

Figure Lengend Snippet: A-B. Representative confocal images showing CAVIN2 (white), Collagen IV (green), TH (orange) expression in the SN (A) and frontal cortex (B) of human postmortem samples from controls and PD donors. C. Quantification of CAVIN2 signal intensity in collagen IV-positive blood vessels associated with acquisitions A and B. The violin plot shows the median (dashed line) and interquartile range (solid line). Data are from five patients and five control donors (n˃ 1,510 vessels analyzed for each condition). Statistical analysis was performed using two-way ANOVA with Tukey’s multiple comparisons correction. D. UMAPs represent CAVIN2 expression level in endothelial nuclei related to Smajić et al. and Kamath et al. E. Graph representing the proportion of CAVIN2 -expressing endothelial nuclei by snRNA-seq (blue bars), and CAVIN2-positive vascular fragments by immunofluorescence (purple bars). F-G. Representative confocal images showing the presence of CAVIN2 (red), Collagen IV (green), TH (white) in blood vessels of the mouse SN (F) , cortex and hippocampus (G ). White arrows point to CAVIN2-positive blood vessels. ( H ) Representative confocal images showing the presence of CAVIN2 (white) and COLLAGEN IV in mouse brain capillaries. Abbreviations: Ctl, control; PD, Parkinson’s disease.

Article Snippet: The second CAVIN2 (Proteintech, 12339-1-AP) and ANXA1 (Invitrogen, 71-3400) antibodies provided similar staining patterns.

Techniques: Expressing, Control, Immunofluorescence

Journal: Frontiers in Immunology

Article Title: Annexin-A1 Tripeptide Attenuates Surgery-Induced Neuroinflammation and Memory Deficits Through Regulation the NLRP3 Inflammasome

doi: 10.3389/fimmu.2022.856254

Figure Lengend Snippet: Decreased ANXA1 expression and induction of the NLRP3 inflammasome in adult mice after orthopedic surgery. Surgery reduced the expression of endogenous ANXA1 in hippocampal homogenate in a time-dependent manner, with the lowest expression found at 24-h (A) . Hence, we treated mice with ANXA1sp (1 mg/kg, i.p.) 30 min before surgery and evaluated NLPR3 inflammasome activation at this timepoint. Postoperative NLRP3 inflammasome activation was significantly attenuated by ANXA1sp (B, C) , as evidenced by Western blot analysis of ASC (D) , cleaved caspase-1 (E) , and cleaved IL-1 (F) in hippocampal homogenate. Surgery-induced hippocampal ATP was also reduced by ANXA1sp treatment (G) . Data are presented as mean ± SEM. n = 4/group. * P < 0.05, ** P < 0.01, *** P < 0.001; analyzed by one-way ANOVA with Bonferroni test. Representative data are from two independent experiments.

Article Snippet: After blocking with 5% nonfat dry milk (170-6404; Bio-Rad), the blot was probed with the following antibodies (1:1000): NLRP3 (LS-C374964; LSBio), ASC (67824S; Cell Signaling), Caspase1 (Cleaved Asp210) (LS-C380449; LSBio), IL-1β (LS-C104813; LSBio), and Annexin-A1 (ANXA1) (3299S; Cell Signaling) or (71-3400; Invitrogen).

Techniques: Expressing, Activation Assay, Western Blot

Journal: Frontiers in Immunology

Article Title: Annexin-A1 Tripeptide Attenuates Surgery-Induced Neuroinflammation and Memory Deficits Through Regulation the NLRP3 Inflammasome

doi: 10.3389/fimmu.2022.856254

Figure Lengend Snippet: Proposed NLPR3 signaling mechanism in postoperative neuroinflammation. Aseptic surgical trauma induces a cytokine storm that engages several pattern recognition receptors (PRRs), including the interlukin-1 (IL-1) receptor. Downstream signaling from these receptors relies on adaptor proteins such as MyD88, which eventually activates NF-κB to perpetuate the inflammatory response. Parallel to this process, surgery also elevated levels of ATP in the hippocampus. In combination with NF-κB activation this induced the NLRP3 inflammasome and ASC specks formation in microglia, thus contributing to postoperative neuroinflammation. Surgery also impaired the expression of hippocampal Annexin-A1 (ANXA1) that is critically involved in inflammation-resolution and can be regulated through our biomimetic peptide. Within this framework we found that treatment with ANXA1sp regulated NLRP3 expression and levels of IL-1β postoperatively. Given the established elevation of IL-1β in preclinical and clinical biofluids of subjects with postoperative neurocognitive disorders, harnessing this pathway may provide safer strategies to treat ensuing neuroinflammation.

Article Snippet: After blocking with 5% nonfat dry milk (170-6404; Bio-Rad), the blot was probed with the following antibodies (1:1000): NLRP3 (LS-C374964; LSBio), ASC (67824S; Cell Signaling), Caspase1 (Cleaved Asp210) (LS-C380449; LSBio), IL-1β (LS-C104813; LSBio), and Annexin-A1 (ANXA1) (3299S; Cell Signaling) or (71-3400; Invitrogen).

Techniques: Activation Assay, Expressing