polyclonal goat anti human endoglin antibody  (R&D Systems)

Journal: Frontiers in Medicine

Article Title: Endoglin and squamous cell carcinomas

doi: 10.3389/fmed.2023.1112573

Figure Lengend Snippet: Six marker FFPE panel for imaging mass spectrometry (Hyperion).

Article Snippet: Antigen retrieval was performed by boiling the sections in a 0.01 M citrate solution (pH 6.0) for 10 min. After being washed with phosphate-buffered saline (PBS), the sections were incubated with polyclonal goat anti-human endoglin (1:400—BAF1097, R&D systems, MN, United States) in PBS/1% bovine serum albumin (BSA), and left overnight at room temperature.

Techniques: Marker, Imaging, Mass Spectrometry

Journal: Frontiers in Medicine

Article Title: Endoglin and squamous cell carcinomas

doi: 10.3389/fmed.2023.1112573

Figure Lengend Snippet: Analysis of squamous cell carcinoma (SCC) primary tumors via immunohistochemistry, all tissues were stained for endoglin expression (brown). The black arrows indicate endothelial endoglin expression. The white arrows indicate epithelial endoglin expression. (A) Representative images of ESCC ( n = 9). (B) Representative images of HNSCC ( n = 5). (C) Representative images of VSCC ( n = 7). Images taken at 100x (left) and 200x (right).

Article Snippet: Antigen retrieval was performed by boiling the sections in a 0.01 M citrate solution (pH 6.0) for 10 min. After being washed with phosphate-buffered saline (PBS), the sections were incubated with polyclonal goat anti-human endoglin (1:400—BAF1097, R&D systems, MN, United States) in PBS/1% bovine serum albumin (BSA), and left overnight at room temperature.

Techniques: Immunohistochemistry, Staining, Expressing

Journal: Frontiers in Medicine

Article Title: Endoglin and squamous cell carcinomas

doi: 10.3389/fmed.2023.1112573

Figure Lengend Snippet: Analysis of HNSCC via imaging mass spectrometry (Hyperion) with a six marker panels. (A) Six images, each depicting the expression of the corresponding marker. (B) A merged image combining pan-cytokeratin, endoglin, and p53 expression. The white arrow indicates cells that co-express pan-cytokeratin and endoglin, which are negative for p53. (C) A merged image combining pan-cytokeratin, endoglin, and CD68 expression. The white arrow indicates cells that co-express pan-cytokeratin and endoglin, which are negative for CD68.

Article Snippet: Antigen retrieval was performed by boiling the sections in a 0.01 M citrate solution (pH 6.0) for 10 min. After being washed with phosphate-buffered saline (PBS), the sections were incubated with polyclonal goat anti-human endoglin (1:400—BAF1097, R&D systems, MN, United States) in PBS/1% bovine serum albumin (BSA), and left overnight at room temperature.

Techniques: Imaging, Mass Spectrometry, Marker, Expressing

Journal: Frontiers in Medicine

Article Title: Endoglin and squamous cell carcinomas

doi: 10.3389/fmed.2023.1112573

Figure Lengend Snippet: Analysis of ESCC via imaging mass spectrometry (Hyperion) with a six marker panels. (A) Six images, each depicting the expression of the corresponding marker. (B) A merged image combining pan-cytokeratin, endoglin, and p53 expression. The white arrows indicate cells that co-express pan-cytokeratin and endoglin, which are negative for p53. (C) A merged image combining pan-cytokeratin, endoglin, and CD68 expression. The white arrow indicates cells that co-express pan-cytokeratin and endoglin, which are negative for CD68. The blue arrow indicates cells that co-express pan-cytokeratin, endoglin, and CD68.

Article Snippet: Antigen retrieval was performed by boiling the sections in a 0.01 M citrate solution (pH 6.0) for 10 min. After being washed with phosphate-buffered saline (PBS), the sections were incubated with polyclonal goat anti-human endoglin (1:400—BAF1097, R&D systems, MN, United States) in PBS/1% bovine serum albumin (BSA), and left overnight at room temperature.

Techniques: Imaging, Mass Spectrometry, Marker, Expressing

Journal: Frontiers in Medicine

Article Title: Endoglin and squamous cell carcinomas

doi: 10.3389/fmed.2023.1112573

Figure Lengend Snippet: Analysis of VSCC via imaging mass spectrometry (Hyperion) with a six marker panel. (A) Six images, each depicting the expression of the corresponding marker. (B) A merged image combining pan-cytokeratin, endoglin, and p53 expression. The white arrows indicate cells that co-express pan-cytokeratin and endoglin. (C) A merged image combining pan-cytokeratin, endoglin, and CD68 expression. The white arrow indicates cells that co-express pan-cytokeratin and endoglin, which are negative for CD68. The blue arrow indicates cells that co-express pan-cytokeratin, endoglin, and CD68.

Article Snippet: Antigen retrieval was performed by boiling the sections in a 0.01 M citrate solution (pH 6.0) for 10 min. After being washed with phosphate-buffered saline (PBS), the sections were incubated with polyclonal goat anti-human endoglin (1:400—BAF1097, R&D systems, MN, United States) in PBS/1% bovine serum albumin (BSA), and left overnight at room temperature.

Techniques: Imaging, Mass Spectrometry, Marker, Expressing

Journal: Frontiers in Medicine

Article Title: Endoglin and squamous cell carcinomas

doi: 10.3389/fmed.2023.1112573

Figure Lengend Snippet: The expression of endoglin by SCC cell lines. Expression by 10 ESCC cell lines, where endoglin expression by TE01 ( p < 0.0001) and TE15 ( p < 0.003) significantly differ from all other cell lines (A) , OSC-19 and FaDu show significantly different endoglin expression ( p = 0.0002) (B) , as is also detected in the three VSCC cell lines (* p = 0.0123, ** p = 0.0025, *** p = 0.0001) with different morphologies (conventional—VC415-C and VC704; spindle—VC415-S) (C) . Endoglin protein levels were determined via western blot (D) and ELISA. Endoglin protein expression by TE01 significantly differs from all other cell lines—( p ≤ 0.0018) (E) . Image for western blot analysis is a representative image of n = 2–3 independent experiments.

Article Snippet: Antigen retrieval was performed by boiling the sections in a 0.01 M citrate solution (pH 6.0) for 10 min. After being washed with phosphate-buffered saline (PBS), the sections were incubated with polyclonal goat anti-human endoglin (1:400—BAF1097, R&D systems, MN, United States) in PBS/1% bovine serum albumin (BSA), and left overnight at room temperature.

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Medicine

Article Title: Endoglin and squamous cell carcinomas

doi: 10.3389/fmed.2023.1112573

Figure Lengend Snippet: Analysis of ALK expression by SCC cell lines, determined via qPCR. Gene expression for ALK1—ALK7 by TE10—endoglin low (A) , TE11—endoglin low (B) , TE01—endoglin high (C) , OSC-19—endoglin positive (D) , FaDu–endoglin high (E) , VC415-C—endoglin low, and VC415-S—endoglin high; *** p = 0.000581 (F) . The effect of endoglin overexpression (OE) on ALK expression was analyzed for TE10; *** p =0.000126; ** p =0.003291 (G) and TE11 (H) , as well as the effect of endoglin knockout (KO) in TE01 (I) .

Article Snippet: Antigen retrieval was performed by boiling the sections in a 0.01 M citrate solution (pH 6.0) for 10 min. After being washed with phosphate-buffered saline (PBS), the sections were incubated with polyclonal goat anti-human endoglin (1:400—BAF1097, R&D systems, MN, United States) in PBS/1% bovine serum albumin (BSA), and left overnight at room temperature.

Techniques: Expressing, Gene Expression, Over Expression, Knock-Out

Journal: Frontiers in Medicine

Article Title: Endoglin and squamous cell carcinomas

doi: 10.3389/fmed.2023.1112573

Figure Lengend Snippet: SCC cells were stimulated with either BMP-6 (TE10 and TE11 only), BMP-9 or TGF-β and the level of phosphorylated SMAD1 and SMAD2 (pSMAD1 and pSMAD2) was determined via western blot (A–C) . The amount of soluble endoglin in the medium of TE10 and TE11 was determined via ELISA (D,E) . Stimulation of OSC-19 (F) , FaDu (G) , VC415-C (H) , and VC415-S (I) with BMP-9/TGF-β/TRC105 was performed, and the levels of pSMAD1 and pSMAD2 were determined via western blot. Western blot images are representative of n = 2–3 per experiment.

Article Snippet: Antigen retrieval was performed by boiling the sections in a 0.01 M citrate solution (pH 6.0) for 10 min. After being washed with phosphate-buffered saline (PBS), the sections were incubated with polyclonal goat anti-human endoglin (1:400—BAF1097, R&D systems, MN, United States) in PBS/1% bovine serum albumin (BSA), and left overnight at room temperature.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Medicine

Article Title: Endoglin and squamous cell carcinomas

doi: 10.3389/fmed.2023.1112573

Figure Lengend Snippet: SCC cell lines were stimulated with BMP-9 or TGF-β and the proliferation of TE10 (A) , TE11 (C) , TE01 (E) , and VC415-S (G) cells was measured via a MTS assay. To assess the effects of endoglin on cell proliferation, MTS assays were performed on endoglin overexpressing (OE) TE10 (B) and TE11 (D) cells. The effects of endoglin knockout (KO) in TE01 (F) and endoglin knockdown (KD) in VC415-S (H) were also assessed via MTS. n = 2–3 for each experiment.

Article Snippet: Antigen retrieval was performed by boiling the sections in a 0.01 M citrate solution (pH 6.0) for 10 min. After being washed with phosphate-buffered saline (PBS), the sections were incubated with polyclonal goat anti-human endoglin (1:400—BAF1097, R&D systems, MN, United States) in PBS/1% bovine serum albumin (BSA), and left overnight at room temperature.

Techniques: MTS Assay, Knock-Out, Knockdown

Journal: Frontiers in Medicine

Article Title: Endoglin and squamous cell carcinomas

doi: 10.3389/fmed.2023.1112573

Figure Lengend Snippet: SCC cell lines were stimulated with BMP-9 or TGF-β and the migration of TE10 (A) , TE11 (C) , TE01 (E) , VC415-S (G) , OSC-19 (I) , and FaDu (J) cells was measured via a wound healing assay. To assess the effects of endoglin on cell migration, wound healing assays were performed on endoglin overexpressing (OE) TE10 (B) and TE11 (D) cells. The effects of endoglin knockout (KO) in TE01 (F) and endoglin knockdown (KD) in VC415-S (H) were also assessed. Finally, the effects of TRC105 on cell migration was assessed in OSC-19 (I) and FaDu cells (J) . n = 2–3 for each experiment.

Article Snippet: Antigen retrieval was performed by boiling the sections in a 0.01 M citrate solution (pH 6.0) for 10 min. After being washed with phosphate-buffered saline (PBS), the sections were incubated with polyclonal goat anti-human endoglin (1:400—BAF1097, R&D systems, MN, United States) in PBS/1% bovine serum albumin (BSA), and left overnight at room temperature.

Techniques: Migration, Wound Healing Assay, Knock-Out, Knockdown

Journal: Frontiers in Medicine

Article Title: Endoglin and squamous cell carcinomas

doi: 10.3389/fmed.2023.1112573

Figure Lengend Snippet: Summary table of the relationship between (altered) endoglin expression, BMP-9 signaling, TGF-β signaling, SCC cell migration, and SCC cell proliferation.

Article Snippet: Antigen retrieval was performed by boiling the sections in a 0.01 M citrate solution (pH 6.0) for 10 min. After being washed with phosphate-buffered saline (PBS), the sections were incubated with polyclonal goat anti-human endoglin (1:400—BAF1097, R&D systems, MN, United States) in PBS/1% bovine serum albumin (BSA), and left overnight at room temperature.

Techniques: Expressing, Migration

Journal: EBioMedicine

Article Title: Reversing pathology in a preclinical model of Alzheimer's disease by hacking cerebrovascular neoangiogenesis with advanced cancer therapeutics.

doi: 10.1016/j.ebiom.2021.103503

Figure Lengend Snippet: Fig. 3. Expression of Ab, the angiogenic marker CD105 and the tight junction protein, ZO1 in aged WT and Tg2576 mice. Whole-brain homogenates from perfused mice were used for molecular analysis of the different biomarkers by immunoblotting. a) Expression of the angiogenesis marker CD105 was significantly higher in the brains of vehicle-treated Tg2576 mice than in the brains of Axitinib-treated Tg2576 or WT animals. b) The expression of tight junction protein, ZO1, was significantly lower in the vehicle-treated Tg2576 mice when compared to the WT mice or Axitinib-treated Tg2576 mice. c) The presence of Ab in the Tg2576 mouse brain was much higher in the vehicle-treated than in Axitinib- treated animals, resembling background levels (e.g. those observed in WT brains). Representative immunoblots are shown for mice from different groups. The anti-Ab antibody used here was initially made against human amyloid but cross-reacts with mouse amyloid under reducing conditions. The data in the histograms are representative of means of individual animals with error bars representing the standard deviation from the mean observed in three separate experiments with WT n= 6 and Tg2576 n= 6 per treatment group. A two-tailed unpaired t-test with was used to calculate significance (* p<0.03; ** p<0.002; *** p<0.0002; **** p<0.0001).

Article Snippet: Antibodies used for immunoblots, immunofluorescence and immunoprecipitation (concentrations were used according to company datasheets unless indicated otherwise): Goat anti-human endoglin / CD105 polyclonal antibody (RRID: AB_354598 R&D systems; AF1097 https://resources.rndsystems.com/pdfs/datasheets/af1097. pdf?v=20210622&_ga=2.123831427.868075561.16243931411326343303.1619741954), anti-b-Amyloid, 1-16 clone 6E-10 (BioLegend; 39320 https://www.biolegend.com/en-us/global-elements/ pdf-popup/purified-anti-beta-amyloid-1-16-antibody-11228?filename=Purified%20anti-beta-Amyloid%201-16%20%20Antibody. pdf&pdfgen=true), Polyclonal Antibody Z0-1 (RRID: AB_2533938 ThermoFisher Scientific; cat. # 61-7300 https://www.thermofisher. com/order/genome-database/dataSheetPdf?producttype=antibody& productsubtype=antibody_primary&productId=61-7300&ver sion=156), Recombinant anti-GAPDH antibody [EPR16891] (RRID: AB_2630358 abcam; ab181602), anti-Actin (C-11) antibody (RRID: AB_630835 Santa Cruz Biotechnology; sc1615 https://search.cosmo bio.co.jp/cosmo_search_p/search_gate2/docs/SCB_/ SC1615.20070227.pdf), rabbit anti-human occludin polyclonal antibody (RRID: AB_881773 abcam; ab31721), anti-CD31 antibody (RRID: AB_726362 abcam; ab28364), anti-mouse serum albumin antibody (RRID: AB_777886 abcam; ab19194), anti-amyloid (1-16) antibody 6E10 (Biolegend; cat. # 803002 https://www.biolegend. com/en-us/global-elements/pdf-popup/purified-anti-beta-amyloid1-16-antibody-11228?filename=Purified%20anti-beta-Amyloid%20116%20%20Antibody.pdf&pdfgen=true), beta Amyloid 1-42 anti-amyloid antibody (RRID: AB_867645 abcam; ab39377).

Techniques: Expressing, Marker, Western Blot, Standard Deviation, Two Tailed Test

Journal: EBioMedicine

Article Title: Reversing pathology in a preclinical model of Alzheimer's disease by hacking cerebrovascular neoangiogenesis with advanced cancer therapeutics.

doi: 10.1016/j.ebiom.2021.103503

Figure Lengend Snippet: Fig. 4. Multiplexed immunofluorescence analysis of WT and Tg2576 mouse brain sec- tions before and after Axitinib treatment. a) Brain sections of mice from different groups were stained for the combination of markers CD105, Ab and CD31. The micro- graph panels are representative of the cortical region of the brains of the mice belong- ing to the different treatment groups. The white grid is representative of the 3D volume of the field of view. The micrographs show the voxels of the field of view of the imaged sections represented by the grid. The data are represented as the mean§ stan- dard deviation. Statistical analysis was performed using unpaired Student’s t-test with the same area imaged from n= 4 to 6 animals per treatment group. (* p<0.03; ** p<0.002; *** p<0.0002; **** p<0.0001). Ab (red); CD31 (blue); CD105 (green), and where coincident staining occurs, combinations are as follows: Red + Green = Yellow; Red + Blue = Purple; Blue + Green =Cyan; Blue + Red + Green =White. The dimensions of each micrograph are 100 X100 X 20 mm. (Scale: each white box = 10 mm) b) Brain sections of mice from different groups were stained for the combination of markers CD105, CD31 and the tight junction protein, Occludin. The micrograph panels are rep- resentative of the cortical region of the brain. CD31 (red); CD105 (green); Occludin (blue) and where coincident staining occurs, combinations are as follows: Red + Green = Yellow; Red + Blue = Purple; Blue + Green =Cyan; Blue + Red + Green =White. Size of micrograph: 100 mm X 100 mm. The data are represented as the mean§ standard devi- ation. Statistical analysis was performed using two-tailed unpaired Student’s t-test with the same area imaged from n= 3 animals per treatment group. (* p<0.03; ** p<0.002; *** p<0.0002; **** p<0.0001). (For interpretation of the references to col- our in this figure, the reader is referred to the web version of this article.)

Article Snippet: Antibodies used for immunoblots, immunofluorescence and immunoprecipitation (concentrations were used according to company datasheets unless indicated otherwise): Goat anti-human endoglin / CD105 polyclonal antibody (RRID: AB_354598 R&D systems; AF1097 https://resources.rndsystems.com/pdfs/datasheets/af1097. pdf?v=20210622&_ga=2.123831427.868075561.16243931411326343303.1619741954), anti-b-Amyloid, 1-16 clone 6E-10 (BioLegend; 39320 https://www.biolegend.com/en-us/global-elements/ pdf-popup/purified-anti-beta-amyloid-1-16-antibody-11228?filename=Purified%20anti-beta-Amyloid%201-16%20%20Antibody. pdf&pdfgen=true), Polyclonal Antibody Z0-1 (RRID: AB_2533938 ThermoFisher Scientific; cat. # 61-7300 https://www.thermofisher. com/order/genome-database/dataSheetPdf?producttype=antibody& productsubtype=antibody_primary&productId=61-7300&ver sion=156), Recombinant anti-GAPDH antibody [EPR16891] (RRID: AB_2630358 abcam; ab181602), anti-Actin (C-11) antibody (RRID: AB_630835 Santa Cruz Biotechnology; sc1615 https://search.cosmo bio.co.jp/cosmo_search_p/search_gate2/docs/SCB_/ SC1615.20070227.pdf), rabbit anti-human occludin polyclonal antibody (RRID: AB_881773 abcam; ab31721), anti-CD31 antibody (RRID: AB_726362 abcam; ab28364), anti-mouse serum albumin antibody (RRID: AB_777886 abcam; ab19194), anti-amyloid (1-16) antibody 6E10 (Biolegend; cat. # 803002 https://www.biolegend. com/en-us/global-elements/pdf-popup/purified-anti-beta-amyloid1-16-antibody-11228?filename=Purified%20anti-beta-Amyloid%20116%20%20Antibody.pdf&pdfgen=true), beta Amyloid 1-42 anti-amyloid antibody (RRID: AB_867645 abcam; ab39377).

Techniques: Staining, Two Tailed Test

Journal: EBioMedicine

Article Title: Reversing pathology in a preclinical model of Alzheimer's disease by hacking cerebrovascular neoangiogenesis with advanced cancer therapeutics.

doi: 10.1016/j.ebiom.2021.103503

Figure Lengend Snippet: Fig. 5. Examination of tight junction proteins by immunofluorescence microscopy in the brains of Tg2576 mice and their WT littermates. a) Brains from perfused mice were used for immunofluorescence analysis of the BBB. Cortical brain sections were stained for the combination of markers CD105, CD31 and the tight junction protein, Occludin. Panel a) shows a micrograph of the cortical region of the brain with merged CD31 (red), CD105 (green) and Occludin (blue) staining whereas the other three panels show CD31, CD105 and Occludin where the field of view was enlarged to reveal a more detailed view of the structures. b) The percentage of tight junction protein disruption is shown graphically (WT B6SJL n= 5 and Tg2576 n= 6). The data are representative of three separate experiments (two-tailed unpaired t-test, * p<0.03; ** p<0.002; *** p<0.0002; **** p<0.0001). c) Images of the brain show an intact BBB and a disrupted BBB. Evans Blue staining is normally very low when the BBB is intact. Post-treatment with Evans Blue, the harvested brains from WT mice showed no colouration, while the brains from vehicle-treated Tg2576 mice appeared blue. d) Post-treatment with Evans Blue, the dye was extracted from the brains, minus the olfactory bulbs and the cerebellum. The absorbance of the extracted dye was read with an ELISA plate reader at 620 nm, and the readings were divided by the weight of the brain. An increase in the uptake of Evans Blue in the brain homogenates prepared from the vehicle-treated Tg2576 mice was indicated by the high absorbance of the dye. No difference was observed between the vehicle- and drug-treated WT (B6/SJL) mice. Axitinib treatment of the Tg2576 mice resulted in less Evans Blue staining than in the vehicle-treated ani- mals, thereby indicating restoration of a functioning BBB. This experiment was repeated twice. (two-tailed unpaired t-test, * p<0.03; ** p<0.002; *** p<0.0002; **** p<0.0001). e) The cortical micrographs here show the voxels of the field of view of the imaged sections represented by the grid. The micrographs are representative of brain sections from mice from a separate experiment, where both Tg2576 and WT animals were treated with Axitinib or delivery vehicle alone, followed by staining for albumin. Red indicates immuno- stained albumin that has leaked into the brain and cyan indicates the cell nucleus (stained with DAPI). Axitinib treatment was associated with less albumin leakage into the brain, indicative of a more functional BBB, in contrast to the greater albumin staining in the vehicle-treated Tg2576. f) The quantification of Albumin is shown as the percentage of pixels in each field of view (n=3). The data are represented as the mean§ standard deviation. Statistical analysis was performed using an unpaired two-tailed t-test. (* p<0.03; ** p<0.002; *** p<0.0002; **** p<0.0001).(For interpretation of the references to colour in this figure, the reader is referred to the web version of this article.)

Article Snippet: Antibodies used for immunoblots, immunofluorescence and immunoprecipitation (concentrations were used according to company datasheets unless indicated otherwise): Goat anti-human endoglin / CD105 polyclonal antibody (RRID: AB_354598 R&D systems; AF1097 https://resources.rndsystems.com/pdfs/datasheets/af1097. pdf?v=20210622&_ga=2.123831427.868075561.16243931411326343303.1619741954), anti-b-Amyloid, 1-16 clone 6E-10 (BioLegend; 39320 https://www.biolegend.com/en-us/global-elements/ pdf-popup/purified-anti-beta-amyloid-1-16-antibody-11228?filename=Purified%20anti-beta-Amyloid%201-16%20%20Antibody. pdf&pdfgen=true), Polyclonal Antibody Z0-1 (RRID: AB_2533938 ThermoFisher Scientific; cat. # 61-7300 https://www.thermofisher. com/order/genome-database/dataSheetPdf?producttype=antibody& productsubtype=antibody_primary&productId=61-7300&ver sion=156), Recombinant anti-GAPDH antibody [EPR16891] (RRID: AB_2630358 abcam; ab181602), anti-Actin (C-11) antibody (RRID: AB_630835 Santa Cruz Biotechnology; sc1615 https://search.cosmo bio.co.jp/cosmo_search_p/search_gate2/docs/SCB_/ SC1615.20070227.pdf), rabbit anti-human occludin polyclonal antibody (RRID: AB_881773 abcam; ab31721), anti-CD31 antibody (RRID: AB_726362 abcam; ab28364), anti-mouse serum albumin antibody (RRID: AB_777886 abcam; ab19194), anti-amyloid (1-16) antibody 6E10 (Biolegend; cat. # 803002 https://www.biolegend. com/en-us/global-elements/pdf-popup/purified-anti-beta-amyloid1-16-antibody-11228?filename=Purified%20anti-beta-Amyloid%20116%20%20Antibody.pdf&pdfgen=true), beta Amyloid 1-42 anti-amyloid antibody (RRID: AB_867645 abcam; ab39377).

Techniques: Microscopy, Staining, Disruption, Two Tailed Test, Enzyme-linked Immunosorbent Assay, Functional Assay, Standard Deviation

Journal: EBioMedicine

Article Title: Reversing pathology in a preclinical model of Alzheimer's disease by hacking cerebrovascular neoangiogenesis with advanced cancer therapeutics.

doi: 10.1016/j.ebiom.2021.103503

Figure Lengend Snippet: Fig. 6. Immunofluorescence colocalization of CD105 with Ab and Occludin. a) Micrographs representing the cortical region of Tg2576 mouse brains depicting co-localization of CD105 and Ab. CD105 (green), Ab (red) and merged where Red + Green = Yellow. Size of each micrograph: 10 mm X 100 mm.b) The scatterplot represents the number and intensity of pixels that are plotted the merged figure (a) of Ab (red) versus CD105 (green). The Thresholded Pearson’s Correlation coefficients are calculated from the scatterplots of 12 sepa- rate comparisons of Ab versus CD105 and are shown in the relative frequency histogram, for an average of (r) = +0.633 with a standard deviation of 0.096, p = 0.01 (statistically sig- nificant, one-tail t-test calculated using free statistics calculator (https://www.danielsoper.com/statcalc) ). c) Micrographs representing cortical region of Tg2576 mouse brains depicting cosiglocalization of CD31 (red), CD105 (green), and Occludin (blue). Merged micrographs show Green + Red = Yellow; Red + Blue = Purple; Blue + Green = Cyan; and Red + Blue + Green = White. Size represented in each micrograph: 100 mm X100 mm. d) The scatterplot represents the number and intensity of pixels that are plotted in the merged figure for CD31 (red) vs Occludin (blue). The Thresholded Pearson’s Correlation coefficients are calculated from the scatterplots of 13 separate comparisons of CD31 versus Occludin and shown in the relative frequency histogram, for an average of (r) = +0.552 with a standard deviation of 0.095, p = 0.03 (statistically significant, one-tail t-test; calculated using free statistics calculator (https://www.danielsoper.com/statcalc)). e) The scatterplot represents the number and intensity of pixels that are plotted in the merged figure for CD105 (green) vs Occludin (blue). The Thresholded Pearson’s Correlation coefficients are calculated from the scatterplots of 10 separate comparisons of CD105 versus Occludin and shown in the relative frequency histogram, for an average of (r) = -0.140 with a standard deviation of 0.073, p = 0.34 (one-tail t-test not statistically significant (https://www.danielsoper. com/statcalc). (For interpretation of the references to colour in this figure, the reader is referred to the web version of this article.)

Article Snippet: Antibodies used for immunoblots, immunofluorescence and immunoprecipitation (concentrations were used according to company datasheets unless indicated otherwise): Goat anti-human endoglin / CD105 polyclonal antibody (RRID: AB_354598 R&D systems; AF1097 https://resources.rndsystems.com/pdfs/datasheets/af1097. pdf?v=20210622&_ga=2.123831427.868075561.16243931411326343303.1619741954), anti-b-Amyloid, 1-16 clone 6E-10 (BioLegend; 39320 https://www.biolegend.com/en-us/global-elements/ pdf-popup/purified-anti-beta-amyloid-1-16-antibody-11228?filename=Purified%20anti-beta-Amyloid%201-16%20%20Antibody. pdf&pdfgen=true), Polyclonal Antibody Z0-1 (RRID: AB_2533938 ThermoFisher Scientific; cat. # 61-7300 https://www.thermofisher. com/order/genome-database/dataSheetPdf?producttype=antibody& productsubtype=antibody_primary&productId=61-7300&ver sion=156), Recombinant anti-GAPDH antibody [EPR16891] (RRID: AB_2630358 abcam; ab181602), anti-Actin (C-11) antibody (RRID: AB_630835 Santa Cruz Biotechnology; sc1615 https://search.cosmo bio.co.jp/cosmo_search_p/search_gate2/docs/SCB_/ SC1615.20070227.pdf), rabbit anti-human occludin polyclonal antibody (RRID: AB_881773 abcam; ab31721), anti-CD31 antibody (RRID: AB_726362 abcam; ab28364), anti-mouse serum albumin antibody (RRID: AB_777886 abcam; ab19194), anti-amyloid (1-16) antibody 6E10 (Biolegend; cat. # 803002 https://www.biolegend. com/en-us/global-elements/pdf-popup/purified-anti-beta-amyloid1-16-antibody-11228?filename=Purified%20anti-beta-Amyloid%20116%20%20Antibody.pdf&pdfgen=true), beta Amyloid 1-42 anti-amyloid antibody (RRID: AB_867645 abcam; ab39377).

Techniques: Standard Deviation