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 following antibodies were used: rabbit anti-Annexin 1 (1:200, Invitrogen, 71-3400), rabbit anti-Annexin 1 (1:200, Abcam, ab214486), rabbit anti-SDPR (CAVIN2) (1:200, Bio-Techne, NBP2-57218), rabbit anti-SDPR (CAVIN2) (1:200, Proteintech, 12339-1-AP), rabbit anti-ANO2 (1:200, Bio-Techne, NBP2-92888), rabbit anti-ANO2 (1:200, Abcam, 113443), mouse anti-GFAP (1:500, Novus Biologicals, NBP1-05197).

Techniques: Expressing, Control, Immunofluorescence

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: International immunopharmacology

Article Title: CAVIN2 attenuates ventilator-induced lung injury in rats by MAPK/ERK1/2 signaling pathway.

doi: 10.1016/j.intimp.2024.113669

Figure Lengend Snippet: Fig. 1. The expression of CAVIN2 was down-regulated and autophagy and inflammation were increased in the lung tissue of VILI rats (A, B) The mRNA and protein expression of CAVIN2 in the lung tissue of rats in the HV groups was detected by qRT-PCR and Western blot, with β-actin as the internal control. (C) Observation of the lung tissue with the naked eye. (F, G) Lung tissue HE staining (magnification 200 × and 400 × ) based on observation with a light microscope and lung injury score. (E) Total protein content in BALF. (D) Wet/ dry weight ratio of the lung tissue. (H-M) Protein and mRNA expression of IL- 1β, IL-6, LC3II/I, Beclin1 and P62 were detected in rat tissues by qRT-PCR and Western blot, with β-actin as the internal control. (N, O) Representative immunohistochemistry images of CAVIN2 and LC3II/I expression of lungs (magnification 200 × and 400 × ). *p < 0.05. **p < 0.01.***p < 0.001.****p < 0.0001.

Article Snippet: The total protein of rat lung tissues and cells was extracted using RIPA lysis buffer (Solarbio, Beijing, China), and the protein concentration was measured using BCA protein assay kit (Solarbio, Beijing, China). proteins were separated by SDS-PAGE and transferred to a polyvinylidene fluoride(PVDF) membrane (Millipore, Sigma Aldrich, USA), which was sealed with 5 % skim milk at 25°C for 1 h, and then incubated with specific primary antibody CAVIN2(1:1000, Proteintech, Chicago, USA) or ERK1/2(1:4000, Proteintech, Chicago, USA) or pERK1/2(1:5000, Proteintech, Chicago, USA) or P62(1:1000, Proteintech, Chicago, USA) or beclin1(1:5000, Proteintech, Chicago, USA) or LC3II/I(1:5000, Proteintech, Chicago, USA)or IL-1β(1:1000, Proteintech, Chicago, USA)or IL-6(1:1000, Proteintech, Chicago, USA) overnight in a refrigerator at 4°C. the membrane was then incubated with a secondary antibody (IgG, 1:7500, Proteintech, Chicago, USA) at 25°C for 1 h. Finally, the protein bands were visualized by CLiNX imaging system (Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Staining, Light Microscopy, Immunohistochemistry

Journal: International immunopharmacology

Article Title: CAVIN2 attenuates ventilator-induced lung injury in rats by MAPK/ERK1/2 signaling pathway.

doi: 10.1016/j.intimp.2024.113669

Figure Lengend Snippet: Fig. 2. Overexpression of CAVIN2 inhibits inflammation and autophagy, thereby alleviating lung injury in VILI rats. (A-B) Expression of CAVIN2 in the lung tissue of rats based on qRT-PCR and Western blot, with β-actin as the internal control. (C) Observation of the lung tissue with the naked eye. (E) Total protein content in BALF. (D) Wet/dry weight ratio of the lung tissue. (F, G) Lung tissue HE staining (magnification 200 × and 400 × ) based on observation with a light microscope and lung injury score. (H-M) Protein and mRNA expression of IL-1β, IL-6, LC3II/I, Beclin1 and P62 in rat tissues were detected by qRT-PCR and Western blot, with β-actin as the internal control. (N, O) Representative immunohistochemistry images of CAVIN2 and LC3II/I expres sion of lungs (magnification 200 × and 400 × ). *p < 0.05. **p < 0.01.***p < 0.001.****p < 0.0001.

Article Snippet: The total protein of rat lung tissues and cells was extracted using RIPA lysis buffer (Solarbio, Beijing, China), and the protein concentration was measured using BCA protein assay kit (Solarbio, Beijing, China). proteins were separated by SDS-PAGE and transferred to a polyvinylidene fluoride(PVDF) membrane (Millipore, Sigma Aldrich, USA), which was sealed with 5 % skim milk at 25°C for 1 h, and then incubated with specific primary antibody CAVIN2(1:1000, Proteintech, Chicago, USA) or ERK1/2(1:4000, Proteintech, Chicago, USA) or pERK1/2(1:5000, Proteintech, Chicago, USA) or P62(1:1000, Proteintech, Chicago, USA) or beclin1(1:5000, Proteintech, Chicago, USA) or LC3II/I(1:5000, Proteintech, Chicago, USA)or IL-1β(1:1000, Proteintech, Chicago, USA)or IL-6(1:1000, Proteintech, Chicago, USA) overnight in a refrigerator at 4°C. the membrane was then incubated with a secondary antibody (IgG, 1:7500, Proteintech, Chicago, USA) at 25°C for 1 h. Finally, the protein bands were visualized by CLiNX imaging system (Shanghai, China).

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Western Blot, Control, Staining, Light Microscopy, Immunohistochemistry

Journal: International immunopharmacology

Article Title: CAVIN2 attenuates ventilator-induced lung injury in rats by MAPK/ERK1/2 signaling pathway.

doi: 10.1016/j.intimp.2024.113669

Figure Lengend Snippet: Fig. 3. CAVIN2 inhibited inflammation and autophagy in MS-induced injury cell models. (A, B) The mRNA and protein expression of CAVIN2 in ATII cells under cyclic mechanical stretch was detected by qRT-PCR and Western blot, with β-actin as the internal control. (A-G) Protein and mRNA expression of IL-1β, IL-6, LC3II/I, Beclin1 and P62 in ATII cells were detected by qRT-PCR and Western blot, with β-actin as the internal control. (H) Immunofluorescence microscopy analysis of the distribution of fluorescence labeled LC3II/I protein (red fluorescence indicates LC3II/I and blue denotes DAPI, Scale:20 μm). *p < 0.05. **p < 0.01.***p < 0.001.****p < 0.0001.

Article Snippet: The total protein of rat lung tissues and cells was extracted using RIPA lysis buffer (Solarbio, Beijing, China), and the protein concentration was measured using BCA protein assay kit (Solarbio, Beijing, China). proteins were separated by SDS-PAGE and transferred to a polyvinylidene fluoride(PVDF) membrane (Millipore, Sigma Aldrich, USA), which was sealed with 5 % skim milk at 25°C for 1 h, and then incubated with specific primary antibody CAVIN2(1:1000, Proteintech, Chicago, USA) or ERK1/2(1:4000, Proteintech, Chicago, USA) or pERK1/2(1:5000, Proteintech, Chicago, USA) or P62(1:1000, Proteintech, Chicago, USA) or beclin1(1:5000, Proteintech, Chicago, USA) or LC3II/I(1:5000, Proteintech, Chicago, USA)or IL-1β(1:1000, Proteintech, Chicago, USA)or IL-6(1:1000, Proteintech, Chicago, USA) overnight in a refrigerator at 4°C. the membrane was then incubated with a secondary antibody (IgG, 1:7500, Proteintech, Chicago, USA) at 25°C for 1 h. Finally, the protein bands were visualized by CLiNX imaging system (Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Immunofluorescence, Microscopy, Fluorescence, Labeling

Journal: International immunopharmacology

Article Title: CAVIN2 attenuates ventilator-induced lung injury in rats by MAPK/ERK1/2 signaling pathway.

doi: 10.1016/j.intimp.2024.113669

Figure Lengend Snippet: Fig. 4. CAVIN2 inhibited autophagy and inflammation in ATII cell VILI modes. (A-D) The CAVIN2 expression in the CAVIN2 overexpression and knock down on ATII cells by qRT-PCR and Western blot. (E-K) Protein and mRNA expression of CAVIN2, IL-1β, IL-6, LC3II/I, Beclin1 and P62 in ATII cells overexpressing CAVIN2 were detected by qRT-PCR and Western blot, with β-actin as the internal control. (L-R) Protein and mRNA expression of CAVIN2, IL-1β, IL-6, LC3II/I, Beclin1 and P62 in ATII cells with CAVIN2 knockdown was determined by qRT-PCR and Western blot, with β-actin as the internal control. (S, T) Immunofluorescence microscopy analysis of the distribution of fluores cence labeled LC3 II/I protein in the ATII cells with overexpressing CAVIN2 and CAVIN2 knockdown (red fluorescence indicates LC3II/I and blue denotes DAPI, Scale:20 μm). *p < 0.05. **p < 0.01.***p < 0.001.****p < 0.0001.

Article Snippet: The total protein of rat lung tissues and cells was extracted using RIPA lysis buffer (Solarbio, Beijing, China), and the protein concentration was measured using BCA protein assay kit (Solarbio, Beijing, China). proteins were separated by SDS-PAGE and transferred to a polyvinylidene fluoride(PVDF) membrane (Millipore, Sigma Aldrich, USA), which was sealed with 5 % skim milk at 25°C for 1 h, and then incubated with specific primary antibody CAVIN2(1:1000, Proteintech, Chicago, USA) or ERK1/2(1:4000, Proteintech, Chicago, USA) or pERK1/2(1:5000, Proteintech, Chicago, USA) or P62(1:1000, Proteintech, Chicago, USA) or beclin1(1:5000, Proteintech, Chicago, USA) or LC3II/I(1:5000, Proteintech, Chicago, USA)or IL-1β(1:1000, Proteintech, Chicago, USA)or IL-6(1:1000, Proteintech, Chicago, USA) overnight in a refrigerator at 4°C. the membrane was then incubated with a secondary antibody (IgG, 1:7500, Proteintech, Chicago, USA) at 25°C for 1 h. Finally, the protein bands were visualized by CLiNX imaging system (Shanghai, China).

Techniques: Expressing, Over Expression, Knockdown, Quantitative RT-PCR, Western Blot, Control, Immunofluorescence, Microscopy, Labeling, Fluorescence

Journal: International immunopharmacology

Article Title: CAVIN2 attenuates ventilator-induced lung injury in rats by MAPK/ERK1/2 signaling pathway.

doi: 10.1016/j.intimp.2024.113669

Figure Lengend Snippet: Fig. 7. CAVIN2 alleviates MS-induced injury by inhibiting ERK1/2 phosphorylation (A-F, H) Protein and mRNA expression of CAVIN2, IL-1β, IL-6, LC3II/I, Beclin1 and P62 in ATII cells were detected by qRT-PCR and Western blot, with β-actin as the internal control. (G) Protein expression of ERK1/2 and p-ERK1/2 in ATII cells were detected by Western blot, with β-actin as the internal Control. (I) Immunofluorescence microscopy analyzed the distribution of fluorescence labeled LC3 II/I protein in the ATII cells (red fluorescence indicates LC3II/I and blue denotes DAPI, Scale:20 μm). *p < 0.05. **p < 0.01.***p < 0.001.****p < 0.0001.

Article Snippet: The total protein of rat lung tissues and cells was extracted using RIPA lysis buffer (Solarbio, Beijing, China), and the protein concentration was measured using BCA protein assay kit (Solarbio, Beijing, China). proteins were separated by SDS-PAGE and transferred to a polyvinylidene fluoride(PVDF) membrane (Millipore, Sigma Aldrich, USA), which was sealed with 5 % skim milk at 25°C for 1 h, and then incubated with specific primary antibody CAVIN2(1:1000, Proteintech, Chicago, USA) or ERK1/2(1:4000, Proteintech, Chicago, USA) or pERK1/2(1:5000, Proteintech, Chicago, USA) or P62(1:1000, Proteintech, Chicago, USA) or beclin1(1:5000, Proteintech, Chicago, USA) or LC3II/I(1:5000, Proteintech, Chicago, USA)or IL-1β(1:1000, Proteintech, Chicago, USA)or IL-6(1:1000, Proteintech, Chicago, USA) overnight in a refrigerator at 4°C. the membrane was then incubated with a secondary antibody (IgG, 1:7500, Proteintech, Chicago, USA) at 25°C for 1 h. Finally, the protein bands were visualized by CLiNX imaging system (Shanghai, China).

Techniques: Phospho-proteomics, Expressing, Quantitative RT-PCR, Western Blot, Control, Immunofluorescence, Microscopy, Fluorescence, Labeling