Structured Review

Millipore thioflavin s
Rac1 activity decreases α-SYN accumulation and aggregation in the neuroblastoma cell line BE(2)-M17. a Representative confocal images of α-SYN over-expressing cells induced with 10 μM retinoic acid (RA) and treated with 10 mM sodium butyrate (SB) for 36 h. Cells were transduced with Control-GFP (upper row), RAC1 (WT)-GFP (middle row) and RAC1 (CA)-GFP (bottom row), and co-stained for <t>Thioflavin</t> S (green) and α-SYN (red). Arrows indicate Thioflavin S positive aggregates with amyloidal structure. b Bar graph showing the quantitative analyses of the neuronal soma area (in percentage %) covered by Thioflavin S positive stain in individual cells transduced with (WT)- or (CA) RAC1 or with the corresponding control. N = 14 (EV), N = 25 (WT), and N = 24 (CA), from at least three independent experiments. Data are presented as mean ± SEM. Statistics, *** P
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Images

1) Product Images from "The Small GTPase RAC1/CED-10 Is Essential in Maintaining Dopaminergic Neuron Function and Survival Against α-Synuclein-Induced Toxicity"

Article Title: The Small GTPase RAC1/CED-10 Is Essential in Maintaining Dopaminergic Neuron Function and Survival Against α-Synuclein-Induced Toxicity

Journal: Molecular Neurobiology

doi: 10.1007/s12035-018-0881-7

Rac1 activity decreases α-SYN accumulation and aggregation in the neuroblastoma cell line BE(2)-M17. a Representative confocal images of α-SYN over-expressing cells induced with 10 μM retinoic acid (RA) and treated with 10 mM sodium butyrate (SB) for 36 h. Cells were transduced with Control-GFP (upper row), RAC1 (WT)-GFP (middle row) and RAC1 (CA)-GFP (bottom row), and co-stained for Thioflavin S (green) and α-SYN (red). Arrows indicate Thioflavin S positive aggregates with amyloidal structure. b Bar graph showing the quantitative analyses of the neuronal soma area (in percentage %) covered by Thioflavin S positive stain in individual cells transduced with (WT)- or (CA) RAC1 or with the corresponding control. N = 14 (EV), N = 25 (WT), and N = 24 (CA), from at least three independent experiments. Data are presented as mean ± SEM. Statistics, *** P
Figure Legend Snippet: Rac1 activity decreases α-SYN accumulation and aggregation in the neuroblastoma cell line BE(2)-M17. a Representative confocal images of α-SYN over-expressing cells induced with 10 μM retinoic acid (RA) and treated with 10 mM sodium butyrate (SB) for 36 h. Cells were transduced with Control-GFP (upper row), RAC1 (WT)-GFP (middle row) and RAC1 (CA)-GFP (bottom row), and co-stained for Thioflavin S (green) and α-SYN (red). Arrows indicate Thioflavin S positive aggregates with amyloidal structure. b Bar graph showing the quantitative analyses of the neuronal soma area (in percentage %) covered by Thioflavin S positive stain in individual cells transduced with (WT)- or (CA) RAC1 or with the corresponding control. N = 14 (EV), N = 25 (WT), and N = 24 (CA), from at least three independent experiments. Data are presented as mean ± SEM. Statistics, *** P

Techniques Used: Activity Assay, Expressing, Transduction, Staining

2) Product Images from "A local insult of okadaic acid in wild-type mice induces tau phosphorylation and protein aggregation in anatomically distinct brain regions"

Article Title: A local insult of okadaic acid in wild-type mice induces tau phosphorylation and protein aggregation in anatomically distinct brain regions

Journal: Acta Neuropathologica Communications

doi: 10.1186/s40478-016-0300-0

Insoluble proteins form NFT-like aggregates rapidly after OA injection at both the injection site and distal brain regions. a β-pleated sheet protein structures are identified by thioflavin-S. Intracellular aggregates are observed at both the injection site and distal sites in the somatosensory cortex at 24 h, which persist 7 days post injection. Scale bar = 50 μM. b High magnification of thioflavin-S positive cells in the amygdala and somatosensory cortex. Scale bar = 10 μm. c Coronal sections taken from OA injected MAPT KO show no phospho-tau immunoreactivity. d No thioflavin positive structures are observed in MAPT KO mice with background staining comparable to the wild-type controls at 24 h. Scale bar = 50 μm
Figure Legend Snippet: Insoluble proteins form NFT-like aggregates rapidly after OA injection at both the injection site and distal brain regions. a β-pleated sheet protein structures are identified by thioflavin-S. Intracellular aggregates are observed at both the injection site and distal sites in the somatosensory cortex at 24 h, which persist 7 days post injection. Scale bar = 50 μM. b High magnification of thioflavin-S positive cells in the amygdala and somatosensory cortex. Scale bar = 10 μm. c Coronal sections taken from OA injected MAPT KO show no phospho-tau immunoreactivity. d No thioflavin positive structures are observed in MAPT KO mice with background staining comparable to the wild-type controls at 24 h. Scale bar = 50 μm

Techniques Used: Injection, Mouse Assay, Staining

3) Product Images from "EFHD2 IS A NOVEL AMYLOID PROTEIN ASSOCIATED TO PATHOLOGICAL TAU IN ALZHEIMER'S DISEASE"

Article Title: EFHD2 IS A NOVEL AMYLOID PROTEIN ASSOCIATED TO PATHOLOGICAL TAU IN ALZHEIMER'S DISEASE

Journal: Journal of neurochemistry

doi: 10.1111/jnc.12155

EFhd2 forms amyloid structures in vitro A , Various concentrations of purified, recombinant His EFhd2 WT (7.5, 30, 40μM) were incubated at 37°C with heparin and DTT (dash, dotted, solid line, respectively). After 20 hrs, samples were added Thioflavin-S (ThioS) and intensity was read in a fluorimeter at Exc.440 nm and Em.550 nm. ThioS stain has a high affinity for cross-β containing structures such as amyloids. EFhd2 showed affinity for this stain and an increase in ThioS binding was observed in a concentration-dependent manner. Baseline ThioS signal of buffer with heparin and DTT is shown (gray line) for comparison with EFhd2-containing reactions. B, Recombinant His EFhd2 WT (40μM) can also bind ThioS stain when incubated without heparin. Formation of cross-β structures by His EFhd2 WT was affected upon the addition of calcium, as it shows a reduction in the binding of ThioS (solid line, minus calcium; dashed lines, plus 1mM CaCl 2 ). Baseline ThioS signal of buffers with heparin and DTT minus (gray line) and plus (dotted line) 1mM CaCl 2 are shown for comparison with protein-containing reactions.
Figure Legend Snippet: EFhd2 forms amyloid structures in vitro A , Various concentrations of purified, recombinant His EFhd2 WT (7.5, 30, 40μM) were incubated at 37°C with heparin and DTT (dash, dotted, solid line, respectively). After 20 hrs, samples were added Thioflavin-S (ThioS) and intensity was read in a fluorimeter at Exc.440 nm and Em.550 nm. ThioS stain has a high affinity for cross-β containing structures such as amyloids. EFhd2 showed affinity for this stain and an increase in ThioS binding was observed in a concentration-dependent manner. Baseline ThioS signal of buffer with heparin and DTT is shown (gray line) for comparison with EFhd2-containing reactions. B, Recombinant His EFhd2 WT (40μM) can also bind ThioS stain when incubated without heparin. Formation of cross-β structures by His EFhd2 WT was affected upon the addition of calcium, as it shows a reduction in the binding of ThioS (solid line, minus calcium; dashed lines, plus 1mM CaCl 2 ). Baseline ThioS signal of buffers with heparin and DTT minus (gray line) and plus (dotted line) 1mM CaCl 2 are shown for comparison with protein-containing reactions.

Techniques Used: In Vitro, Purification, Recombinant, Incubation, Staining, Binding Assay, Concentration Assay

4) Product Images from "Defined ?-synuclein prion-like molecular assemblies spreading in cell culture"

Article Title: Defined ?-synuclein prion-like molecular assemblies spreading in cell culture

Journal: BMC Neuroscience

doi: 10.1186/1471-2202-15-69

Thioflavin-S (ThS) positive staining of endogenous α-syn aggregates. SH-SY5Y cells were infected with recombinant human α-syn short amyloid fibrils (SF: short amyloid fibrils of α-syn) and sub-passed at sixth passage (p6). The deposition and level of α-syn (red) in α-syn-infected cell lines after six passages were detected by anti-α-syn antibody (Additional file 6 : Table S2). The colocalization was observed between the ThS signal (yellow) and anti-human α-syn antibody (red). The nuclei (blue) were stained with DAPI. Scale bars, 12 μm.
Figure Legend Snippet: Thioflavin-S (ThS) positive staining of endogenous α-syn aggregates. SH-SY5Y cells were infected with recombinant human α-syn short amyloid fibrils (SF: short amyloid fibrils of α-syn) and sub-passed at sixth passage (p6). The deposition and level of α-syn (red) in α-syn-infected cell lines after six passages were detected by anti-α-syn antibody (Additional file 6 : Table S2). The colocalization was observed between the ThS signal (yellow) and anti-human α-syn antibody (red). The nuclei (blue) were stained with DAPI. Scale bars, 12 μm.

Techniques Used: Staining, Infection, Recombinant

5) Product Images from "Curcuminoid submicron particle ameliorates cognitive deficits and decreases amyloid pathology in Alzheimer’s disease mouse model"

Article Title: Curcuminoid submicron particle ameliorates cognitive deficits and decreases amyloid pathology in Alzheimer’s disease mouse model

Journal: Oncotarget

doi: 10.18632/oncotarget.24369

CSP decreased the amyloid deposition in the hippocampus of APP mice (A) Representative images of β-sheet amyloid plaques in the hippocampus of APP and APP/CSP mice stained by Thioflavin-S. Scale bar = 200 μm. (B) Quantitative analysis of the number of the plaques in the hippocampus. N = 6 mice/group, 6-10 slices per mouse. (C-D) The levels of Aβ42 (C) and total Aβ (D) in the hippocampal lysate were determined by ELISA. (E) Aβ42/total Aβ were unchanged in APP mice treated with CSP. N = 13-17 mice/group. Results were analyzed by t test. ** , p
Figure Legend Snippet: CSP decreased the amyloid deposition in the hippocampus of APP mice (A) Representative images of β-sheet amyloid plaques in the hippocampus of APP and APP/CSP mice stained by Thioflavin-S. Scale bar = 200 μm. (B) Quantitative analysis of the number of the plaques in the hippocampus. N = 6 mice/group, 6-10 slices per mouse. (C-D) The levels of Aβ42 (C) and total Aβ (D) in the hippocampal lysate were determined by ELISA. (E) Aβ42/total Aβ were unchanged in APP mice treated with CSP. N = 13-17 mice/group. Results were analyzed by t test. ** , p

Techniques Used: Mouse Assay, Staining, Enzyme-linked Immunosorbent Assay

6) Product Images from "Otophylloside B Protects Against Aβ Toxicity in Caenorhabditis elegans Models of Alzheimer’s Disease"

Article Title: Otophylloside B Protects Against Aβ Toxicity in Caenorhabditis elegans Models of Alzheimer’s Disease

Journal: Natural Products and Bioprospecting

doi: 10.1007/s13659-017-0122-1

Ot B decreased Aβ deposition by depressing expression of Aβ . a Thioflavin S staining of CL2006. Ot B treated CL2006 worms were stained with thioflavin S on day 3 and day 5. White arrows indicate Aβ deposits. b Number of Aβ deposits in the worm head region. 50 μM Ot B significantly reduced the number of Aβ deposits in CL2006 both at day 3 and day 5. c The transcript level of Aβ , measured by qRT-PCR. The transcript level of Aβ was significantly downregulated by Ot B. The data was normalized to the expression of cdc - 42 . Each bar represents the mean value of three independent experiments with error bars representing SEM. ** represents p
Figure Legend Snippet: Ot B decreased Aβ deposition by depressing expression of Aβ . a Thioflavin S staining of CL2006. Ot B treated CL2006 worms were stained with thioflavin S on day 3 and day 5. White arrows indicate Aβ deposits. b Number of Aβ deposits in the worm head region. 50 μM Ot B significantly reduced the number of Aβ deposits in CL2006 both at day 3 and day 5. c The transcript level of Aβ , measured by qRT-PCR. The transcript level of Aβ was significantly downregulated by Ot B. The data was normalized to the expression of cdc - 42 . Each bar represents the mean value of three independent experiments with error bars representing SEM. ** represents p

Techniques Used: Expressing, Staining, Quantitative RT-PCR

7) Product Images from "Mitochondrial fusion suppresses tau pathology–induced neurodegeneration and cognitive decline"

Article Title: Mitochondrial fusion suppresses tau pathology–induced neurodegeneration and cognitive decline

Journal: Journal of Alzheimer's disease : JAD

doi: 10.3233/JAD-215175

Inhibition of tau hyperphosphorylation and aggregation in P301S mice by neuronal Mfn2. A) Representative immunocytochemistry of phosphorylated tau recognized by PHF1 and AT8 in hippocampal regions of 11-month-old NTG, TMFN, P301S, and P301S/TMFN mice. B, C) Representative immunoblot (B) and quantification (C) of phosphorylated tau (recognized by both PHF1 and AT8) in total hippocampus lysates of 11-month-old NTG, TMFN, P301S, and P301S/TMFN mice ( n = 4 mice per group). D) Representative EM images of paired helical filaments noted in hippocampal neurons of P301S mice. E) Representative fluorescent images for AT8 (red), Thioflavin-S (green), and DAPI (blue) staining in hippocampal neurons of 11-month-old NTG, TMFN, P301S, and P301S/TMFN mice. The data are expressed as the means ± SEM and were analyzed by Student’s t-test. *p
Figure Legend Snippet: Inhibition of tau hyperphosphorylation and aggregation in P301S mice by neuronal Mfn2. A) Representative immunocytochemistry of phosphorylated tau recognized by PHF1 and AT8 in hippocampal regions of 11-month-old NTG, TMFN, P301S, and P301S/TMFN mice. B, C) Representative immunoblot (B) and quantification (C) of phosphorylated tau (recognized by both PHF1 and AT8) in total hippocampus lysates of 11-month-old NTG, TMFN, P301S, and P301S/TMFN mice ( n = 4 mice per group). D) Representative EM images of paired helical filaments noted in hippocampal neurons of P301S mice. E) Representative fluorescent images for AT8 (red), Thioflavin-S (green), and DAPI (blue) staining in hippocampal neurons of 11-month-old NTG, TMFN, P301S, and P301S/TMFN mice. The data are expressed as the means ± SEM and were analyzed by Student’s t-test. *p

Techniques Used: Inhibition, Mouse Assay, Immunocytochemistry, Staining

8) Product Images from "Erinacine A-enriched Hericium erinaceus mycelium ameliorates Alzheimer’s disease-related pathologies in APPswe/PS1dE9 transgenic mice"

Article Title: Erinacine A-enriched Hericium erinaceus mycelium ameliorates Alzheimer’s disease-related pathologies in APPswe/PS1dE9 transgenic mice

Journal: Journal of Biomedical Science

doi: 10.1186/s12929-016-0266-z

HE-Et and HE-My reduce amyloid plaque burden in the area include the cerebral cortex and hippocampus of APP/PS1 mice. Five month-old APP/PS1 mice were orally administered with vehicle (Veh, n = 6), HE-Et ( n = 5) and HE-My ( n = 6) for 30 days. A . The representative fluorescent images of amyloid plaques detected by thioflavin S (ThS) staining ( white in a, e and i, and green in c, d, g, h, k and l) and immunohistochemical staining with AB-10 antibody (white in b, f and j, red in c, d, g, h, k and l) in the area including parietal cortex and hippocampus. Sale bar: 500 μm. A typical plaque is magnified and shows in the right side of each image. Sale bar: 20 μm. B . shows both the plaque number and burden in ThS- and AB-10-stained semi-cerebral sphere calculated by image analysis software. Plaque burden is displayed as a percentage of the area occupied by ThS- or AB-10-stained signal in the full area of interest. C . The structure of amyloid plaque in the cerebral cortex of APP/PS1 mice. The amyloid plaque in cerebral cortex of 6 months-old APP/PS1 mice was detected by ThS-staining ( white in a and d, and green in c and f) and immunohistochemical staining with AB-10 antibody ( white in b and e, and red in c and f). Upper panels show the representative Z-projection (3 dimensions, XYZ, A-C) of a typical plaque. Lower panels show the 2 dimensional images (XY, XZ, and YZ, D-F). The image indicated by arrow indicated the putative plaque unit. Sale bar: 10 μm. Single fluorescent images were presented as grayscale to enhance resolution
Figure Legend Snippet: HE-Et and HE-My reduce amyloid plaque burden in the area include the cerebral cortex and hippocampus of APP/PS1 mice. Five month-old APP/PS1 mice were orally administered with vehicle (Veh, n = 6), HE-Et ( n = 5) and HE-My ( n = 6) for 30 days. A . The representative fluorescent images of amyloid plaques detected by thioflavin S (ThS) staining ( white in a, e and i, and green in c, d, g, h, k and l) and immunohistochemical staining with AB-10 antibody (white in b, f and j, red in c, d, g, h, k and l) in the area including parietal cortex and hippocampus. Sale bar: 500 μm. A typical plaque is magnified and shows in the right side of each image. Sale bar: 20 μm. B . shows both the plaque number and burden in ThS- and AB-10-stained semi-cerebral sphere calculated by image analysis software. Plaque burden is displayed as a percentage of the area occupied by ThS- or AB-10-stained signal in the full area of interest. C . The structure of amyloid plaque in the cerebral cortex of APP/PS1 mice. The amyloid plaque in cerebral cortex of 6 months-old APP/PS1 mice was detected by ThS-staining ( white in a and d, and green in c and f) and immunohistochemical staining with AB-10 antibody ( white in b and e, and red in c and f). Upper panels show the representative Z-projection (3 dimensions, XYZ, A-C) of a typical plaque. Lower panels show the 2 dimensional images (XY, XZ, and YZ, D-F). The image indicated by arrow indicated the putative plaque unit. Sale bar: 10 μm. Single fluorescent images were presented as grayscale to enhance resolution

Techniques Used: Mouse Assay, Staining, Immunohistochemistry, Software

9) Product Images from "Late-onset Parkinsonism in NF?B/c-Rel-deficient mice"

Article Title: Late-onset Parkinsonism in NF?B/c-Rel-deficient mice

Journal: Brain

doi: 10.1093/brain/aws193

Accumulation of α-synuclein in 18-month-old c-rel −/− and wild-type mice. ( A and D ) Representative photomicrographs showing tyrosine hydroxylase (blue) and α-synuclein (brown) double staining in the substantia nigra of 18-month-old wild-type ( A and C ) and c-rel −/− mice ( B and D ). Note that the increase of brown α-synuclein staining in the c-rel −/− mice ( d ) completely covered the blue tyrosine hydroxylase immunoreactivity observed in wild-type mice ( c ). ( E - L ) Representative photomicrographs showing α-synuclein (red), thioflavin S (green) and tyrosine hydroxylase (blue) triple staining in the substatia nigra of c-rel −/− ( E–H ) and wild-type ( I–L ) mice. Note that the α-synuclein-positive aggregates in the brain of c-rel −/− mice were also thioflavin S immunopositive, as visualized in the high-magnification squares in panels E – H . Scale bars: in A = 3 mm for A and B ; in C = 750 µm for C and D ; in insert c = 10 µm for c and d ; in E = 50 µm for E–L ; in insert e = 10 µm for e – h . Panels A–L are representative of three independent experiments ( n = 3 animals per group). ( M ) Representative western blot showing sequential α-synuclein extraction from diverse brain areas in 18-month-old wild-type and c-rel −/− (-/-) mice. Recombinant α-synuclein (+) solution was used as positive control. Note the increase in α-synuclein-immunopositive bands in the mesencephalon of c-rel −/− mice when compared with wild-type animals, as well as the α-synuclein immunopositive band in the urea extracts from c-rel −/− mice. ( N ) Densitometric analysis of α-synuclein-immunopositive bands in the TBS and TBS/Triton fractions. Note the significant increase of α-synuclein in the mesencephalon of c-rel −/− mice. Data represent the mean ± SEM ( n = 3 animals per group, * P
Figure Legend Snippet: Accumulation of α-synuclein in 18-month-old c-rel −/− and wild-type mice. ( A and D ) Representative photomicrographs showing tyrosine hydroxylase (blue) and α-synuclein (brown) double staining in the substantia nigra of 18-month-old wild-type ( A and C ) and c-rel −/− mice ( B and D ). Note that the increase of brown α-synuclein staining in the c-rel −/− mice ( d ) completely covered the blue tyrosine hydroxylase immunoreactivity observed in wild-type mice ( c ). ( E - L ) Representative photomicrographs showing α-synuclein (red), thioflavin S (green) and tyrosine hydroxylase (blue) triple staining in the substatia nigra of c-rel −/− ( E–H ) and wild-type ( I–L ) mice. Note that the α-synuclein-positive aggregates in the brain of c-rel −/− mice were also thioflavin S immunopositive, as visualized in the high-magnification squares in panels E – H . Scale bars: in A = 3 mm for A and B ; in C = 750 µm for C and D ; in insert c = 10 µm for c and d ; in E = 50 µm for E–L ; in insert e = 10 µm for e – h . Panels A–L are representative of three independent experiments ( n = 3 animals per group). ( M ) Representative western blot showing sequential α-synuclein extraction from diverse brain areas in 18-month-old wild-type and c-rel −/− (-/-) mice. Recombinant α-synuclein (+) solution was used as positive control. Note the increase in α-synuclein-immunopositive bands in the mesencephalon of c-rel −/− mice when compared with wild-type animals, as well as the α-synuclein immunopositive band in the urea extracts from c-rel −/− mice. ( N ) Densitometric analysis of α-synuclein-immunopositive bands in the TBS and TBS/Triton fractions. Note the significant increase of α-synuclein in the mesencephalon of c-rel −/− mice. Data represent the mean ± SEM ( n = 3 animals per group, * P

Techniques Used: Mouse Assay, Double Staining, Staining, Western Blot, Recombinant, Positive Control

10) Product Images from "Nicotinamide phosphoribosyltransferase-related signaling pathway in early Alzheimer's disease mouse models"

Article Title: Nicotinamide phosphoribosyltransferase-related signaling pathway in early Alzheimer's disease mouse models

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2019.10782

Histological images of Thioflavin S staining in the cortex and hippocampus and the effect of NAD + and FK866 on amyloid plaques. Scale bar, 100 µm. **P
Figure Legend Snippet: Histological images of Thioflavin S staining in the cortex and hippocampus and the effect of NAD + and FK866 on amyloid plaques. Scale bar, 100 µm. **P

Techniques Used: Staining

11) Product Images from "Epigenetic mechanisms underlying cognitive impairment and Alzheimer disease hallmarks in 5XFAD mice"

Article Title: Epigenetic mechanisms underlying cognitive impairment and Alzheimer disease hallmarks in 5XFAD mice

Journal: Aging (Albany NY)

doi: 10.18632/aging.100906

Histological images of amyloid plaques stained with thioflavin-S in female mice aged 2 and 8 months (Wt and 5XFAD). There is a heavy load of plaques in the majority of the brain areas ilustrated in 5XFAD the brain section ( A ). Representative Western blot for NeuN (B, C) and Synaptophysin ( D, E ). Bars represent mean ± Standard Error of the Mean (SEM), n = 4 for each group; DG: Dentate Gyrus. Scale bar for histochemical images is indicated in the Picture; *p
Figure Legend Snippet: Histological images of amyloid plaques stained with thioflavin-S in female mice aged 2 and 8 months (Wt and 5XFAD). There is a heavy load of plaques in the majority of the brain areas ilustrated in 5XFAD the brain section ( A ). Representative Western blot for NeuN (B, C) and Synaptophysin ( D, E ). Bars represent mean ± Standard Error of the Mean (SEM), n = 4 for each group; DG: Dentate Gyrus. Scale bar for histochemical images is indicated in the Picture; *p

Techniques Used: Staining, Mouse Assay, Western Blot

12) Product Images from "Curcuminoid submicron particle ameliorates cognitive deficits and decreases amyloid pathology in Alzheimer’s disease mouse model"

Article Title: Curcuminoid submicron particle ameliorates cognitive deficits and decreases amyloid pathology in Alzheimer’s disease mouse model

Journal: Oncotarget

doi: 10.18632/oncotarget.24369

CSP decreased the amyloid deposition in the hippocampus of APP mice (A) Representative images of β-sheet amyloid plaques in the hippocampus of APP and APP/CSP mice stained by Thioflavin-S. Scale bar = 200 μm. (B) Quantitative analysis of the number of the plaques in the hippocampus. N = 6 mice/group, 6-10 slices per mouse. (C-D) The levels of Aβ42 (C) and total Aβ (D) in the hippocampal lysate were determined by ELISA. (E) Aβ42/total Aβ were unchanged in APP mice treated with CSP. N = 13-17 mice/group. Results were analyzed by t test. ** , p
Figure Legend Snippet: CSP decreased the amyloid deposition in the hippocampus of APP mice (A) Representative images of β-sheet amyloid plaques in the hippocampus of APP and APP/CSP mice stained by Thioflavin-S. Scale bar = 200 μm. (B) Quantitative analysis of the number of the plaques in the hippocampus. N = 6 mice/group, 6-10 slices per mouse. (C-D) The levels of Aβ42 (C) and total Aβ (D) in the hippocampal lysate were determined by ELISA. (E) Aβ42/total Aβ were unchanged in APP mice treated with CSP. N = 13-17 mice/group. Results were analyzed by t test. ** , p

Techniques Used: Mouse Assay, Staining, Enzyme-linked Immunosorbent Assay

13) Product Images from "Deletion of CD14 attenuates AD pathology by influencing the brain's inflammatory milieu"

Article Title: Deletion of CD14 attenuates AD pathology by influencing the brain's inflammatory milieu

Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

doi: 10.1523/JNEUROSCI.2637-10.2010

Deletion of CD14 reduces Aβ plaque burden Cortical 6E10 immunohistochemistry of TgCD14 +/+ ( A ) and TgCD14 −/− ( B ) mice. Scale bar = 400 µm. Cortical Thioflavin S immunohistochemistry on TgCD14 +/+ ( C ) and Tg + CD14 −/− ( D ) mice. Scale bar = 100 µm.
Figure Legend Snippet: Deletion of CD14 reduces Aβ plaque burden Cortical 6E10 immunohistochemistry of TgCD14 +/+ ( A ) and TgCD14 −/− ( B ) mice. Scale bar = 400 µm. Cortical Thioflavin S immunohistochemistry on TgCD14 +/+ ( C ) and Tg + CD14 −/− ( D ) mice. Scale bar = 100 µm.

Techniques Used: Immunohistochemistry, Mouse Assay

14) Product Images from "A nontransgenic mouse model shows inducible amyloid-? (A?) peptide deposition and elucidates the role of apolipoprotein E in the amyloid cascade"

Article Title: A nontransgenic mouse model shows inducible amyloid-? (A?) peptide deposition and elucidates the role of apolipoprotein E in the amyloid cascade

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.0404458101

Quantitative determination of the numbers of Aβ42 ( A ), Aβ40 ( B ), and fibrillar Aβ ( C ) deposits in brains of apoE4-transgenic (ApoE4-tg), apoE3-transgenic (ApoE3-tg), apoE-deficient (ApoE-def.), and control mice after i.c.v. infusion of thiorphan for 1 month (see Materials and Methods ). Coronal sections at the level of maximal Aβ deposition (bregma –2.8) were stained with anti-Aβ42 (mAb G211) or anti-Aβ40 (mAb G210) antibodies or with thioflavin-S (see Materials and Methods ). Values (means ± SD of four or five mice per group) for each of the Aβ deposition measurements are for two sections per mouse. * , P
Figure Legend Snippet: Quantitative determination of the numbers of Aβ42 ( A ), Aβ40 ( B ), and fibrillar Aβ ( C ) deposits in brains of apoE4-transgenic (ApoE4-tg), apoE3-transgenic (ApoE3-tg), apoE-deficient (ApoE-def.), and control mice after i.c.v. infusion of thiorphan for 1 month (see Materials and Methods ). Coronal sections at the level of maximal Aβ deposition (bregma –2.8) were stained with anti-Aβ42 (mAb G211) or anti-Aβ40 (mAb G210) antibodies or with thioflavin-S (see Materials and Methods ). Values (means ± SD of four or five mice per group) for each of the Aβ deposition measurements are for two sections per mouse. * , P

Techniques Used: Transgenic Assay, Mouse Assay, Staining

Rostrocaudal distribution of Aβ42 ( A ), Aβ40 ( B ), and fibrillar Aβ ( C ) deposits in brains of apoE-transgenic and control mice after inhibition of neprilysin. ApoE4-transgenic (•), apoE3-transgenic (○), apoE-deficient (▵), and control (▴) mice were infused i.c.v. with thiorphan for 1 month (see Materials and Methods ). Brains were then cut coronally (–4 mm to +3 mm from bregma), and consecutive coronal sections at the indicated positions were stained for Aβ42 (mAb G211), Aβ40 (mAb G210), or fibrillar Aβ deposits (thioflavin-S) (see Materials and Methods ). Values (means ± SD of four or five mice per group) are for two sections per mouse at each of the indicated bregma levels.
Figure Legend Snippet: Rostrocaudal distribution of Aβ42 ( A ), Aβ40 ( B ), and fibrillar Aβ ( C ) deposits in brains of apoE-transgenic and control mice after inhibition of neprilysin. ApoE4-transgenic (•), apoE3-transgenic (○), apoE-deficient (▵), and control (▴) mice were infused i.c.v. with thiorphan for 1 month (see Materials and Methods ). Brains were then cut coronally (–4 mm to +3 mm from bregma), and consecutive coronal sections at the indicated positions were stained for Aβ42 (mAb G211), Aβ40 (mAb G210), or fibrillar Aβ deposits (thioflavin-S) (see Materials and Methods ). Values (means ± SD of four or five mice per group) are for two sections per mouse at each of the indicated bregma levels.

Techniques Used: Transgenic Assay, Mouse Assay, Inhibition, Staining

Effects of the apoE genotype on Aβ deposition after inhibition of neprilysin. ( A ) Representative micrographs of cortical fields of coronal brain sections (bregma –2.8 mm) from apoE4-(ApoE4-tg.) and apoE3-(ApoE3-tg.) transgenic, apoE-deficient (ApoE-def.), and control mice infused i.c.v. for 1 month with the neprilysin inhibitor thiorphan (see Materials and Methods ). ( B ) Representative immunoblots of soluble (SAβ) and insoluble (InAβ) Aβ of the same groups of three mice each. Immunohistochemistry (anti-Aβ42 mAb G211 and anti Aβ40 mAb G210), histochemistry (thioflavin-S), and immunoblot analysis (anti-Aβ mAb 4G8) were performed as described in Materials and Methods . (Scale bar, 50 μm.)
Figure Legend Snippet: Effects of the apoE genotype on Aβ deposition after inhibition of neprilysin. ( A ) Representative micrographs of cortical fields of coronal brain sections (bregma –2.8 mm) from apoE4-(ApoE4-tg.) and apoE3-(ApoE3-tg.) transgenic, apoE-deficient (ApoE-def.), and control mice infused i.c.v. for 1 month with the neprilysin inhibitor thiorphan (see Materials and Methods ). ( B ) Representative immunoblots of soluble (SAβ) and insoluble (InAβ) Aβ of the same groups of three mice each. Immunohistochemistry (anti-Aβ42 mAb G211 and anti Aβ40 mAb G210), histochemistry (thioflavin-S), and immunoblot analysis (anti-Aβ mAb 4G8) were performed as described in Materials and Methods . (Scale bar, 50 μm.)

Techniques Used: Inhibition, Transgenic Assay, Mouse Assay, Western Blot, Immunohistochemistry

15) Product Images from "MyD88 Deficiency Ameliorates ?-Amyloidosis in an Animal Model of Alzheimer's Disease"

Article Title: MyD88 Deficiency Ameliorates ?-Amyloidosis in an Animal Model of Alzheimer's Disease

Journal: The American Journal of Pathology

doi: 10.1016/j.ajpath.2011.05.045

Detection of diffuse and fibrillar Aβ deposits by anti-Aβ antibody and thioflavin S and quantification of buffer-extractable and buffer-unextractable Aβ by ELISA in APP MyD88 −/− and APP mice. Aβ deposits
Figure Legend Snippet: Detection of diffuse and fibrillar Aβ deposits by anti-Aβ antibody and thioflavin S and quantification of buffer-extractable and buffer-unextractable Aβ by ELISA in APP MyD88 −/− and APP mice. Aβ deposits

Techniques Used: Enzyme-linked Immunosorbent Assay, Mouse Assay

16) Product Images from "(E)-2-Methoxy-4-(3-(4-methoxyphenyl) prop-1-en-1-yl) Phenol Ameliorates LPS-Mediated Memory Impairment by Inhibition of STAT3 Pathway"

Article Title: (E)-2-Methoxy-4-(3-(4-methoxyphenyl) prop-1-en-1-yl) Phenol Ameliorates LPS-Mediated Memory Impairment by Inhibition of STAT3 Pathway

Journal: Neuromolecular Medicine

doi: 10.1007/s12017-017-8469-3

Inhibitory effects of MMPP on accumulation of Aβ 1-42 in the brain of LPS-injected mice. ( a ) Aβ accumulation in the brains of LPS-injected mice was determined by thioflavin S staining. The levels of Aβ 1-42 in mice brain ( n = 5) were measured by ELISA ( b ). The activity of β-secretase in mice brain ( n = 5) was investigated using assay kit ( c ). The expression of APP, BACE1 and C99 was detected by Western blotting using specific antibodies in the mouse brain ( d ). For the cropped images, samples were run in the same gels under same experimental conditions and processed in parallel. Each blot is representative for three experiments
Figure Legend Snippet: Inhibitory effects of MMPP on accumulation of Aβ 1-42 in the brain of LPS-injected mice. ( a ) Aβ accumulation in the brains of LPS-injected mice was determined by thioflavin S staining. The levels of Aβ 1-42 in mice brain ( n = 5) were measured by ELISA ( b ). The activity of β-secretase in mice brain ( n = 5) was investigated using assay kit ( c ). The expression of APP, BACE1 and C99 was detected by Western blotting using specific antibodies in the mouse brain ( d ). For the cropped images, samples were run in the same gels under same experimental conditions and processed in parallel. Each blot is representative for three experiments

Techniques Used: Injection, Mouse Assay, Staining, Enzyme-linked Immunosorbent Assay, Activity Assay, Expressing, Western Blot

17) Product Images from "The Effect of Iron in MR Imaging and Transverse Relaxation of Amyloid-Beta Plaques in Alzheimer’s Disease"

Article Title: The Effect of Iron in MR Imaging and Transverse Relaxation of Amyloid-Beta Plaques in Alzheimer’s Disease

Journal: NMR in biomedicine

doi: 10.1002/nbm.3247

Amyloid-beta staining (a,e), iron deposition (b,f), T 2 *-weighted MR images (c,g), and susceptibility weighted images (d,h) of saline (left) and DFO treated (right) AD tissue samples. Thioflavin-S stained Aβ plaques in saline treated (red arrows
Figure Legend Snippet: Amyloid-beta staining (a,e), iron deposition (b,f), T 2 *-weighted MR images (c,g), and susceptibility weighted images (d,h) of saline (left) and DFO treated (right) AD tissue samples. Thioflavin-S stained Aβ plaques in saline treated (red arrows

Techniques Used: Staining

T 2 *-weighted MR images (a, d), microscopy images of Perl’s stain for iron (b, e), and fluorescent thioflavin-S for Aβ plaques (c, f) of sequentially cut AD tissue samples treated with saline (a, b, and c) and deferoxamine (d, e, and f).
Figure Legend Snippet: T 2 *-weighted MR images (a, d), microscopy images of Perl’s stain for iron (b, e), and fluorescent thioflavin-S for Aβ plaques (c, f) of sequentially cut AD tissue samples treated with saline (a, b, and c) and deferoxamine (d, e, and f).

Techniques Used: Microscopy, Staining

18) Product Images from "Ultrasound-mediated augmented exosome release from astrocytes alleviates amyloid-β-induced neurotoxicity"

Article Title: Ultrasound-mediated augmented exosome release from astrocytes alleviates amyloid-β-induced neurotoxicity

Journal: Theranostics

doi: 10.7150/thno.52436

Brain-targeted delivery of US-HA-Exo combined with FUS-mediated BBB opening. (A, B) Schematic of FUS-BBB opening-assisted delivery of exosomes. (C) Brain sections obtained from 10-month-old APP/PS1 mice were immuno-stained for Aβ, scale bar: 100 µm. (D) The percentage area of positive amyloid-β stainning was quantified. (E, F) Aβ plaques in the brain were detected by thioflavin-S staining (scale bar: 100 µm), and quantified. (G) H E staining of major organs of mice, scale bar: 100 µm.
Figure Legend Snippet: Brain-targeted delivery of US-HA-Exo combined with FUS-mediated BBB opening. (A, B) Schematic of FUS-BBB opening-assisted delivery of exosomes. (C) Brain sections obtained from 10-month-old APP/PS1 mice were immuno-stained for Aβ, scale bar: 100 µm. (D) The percentage area of positive amyloid-β stainning was quantified. (E, F) Aβ plaques in the brain were detected by thioflavin-S staining (scale bar: 100 µm), and quantified. (G) H E staining of major organs of mice, scale bar: 100 µm.

Techniques Used: Mouse Assay, Staining

19) Product Images from "Significant structural but not physiological changes in cortical neurons of 12-month-old Tg2576 mice"

Article Title: Significant structural but not physiological changes in cortical neurons of 12-month-old Tg2576 mice

Journal: Neurobiology of disease

doi: 10.1016/j.nbd.2008.07.014

Representative slice and streptavidin-Alexa labeled cells A . Photomicrograph of a slice from a Tg2576 mouse stained with thioflavin-S showing a low density of fibrillar plaques (arrowheads) in the cortex and hippocampus (inset: boxed area at higher magnification). B . Representative streptavidin-Alexa 488 labeled layer 3 pyramidal cells from Tg2576 (top) and Wt (bottom) mice. Scale bars = A: 1 mm, inset: 25 µm; B: 100 µm.
Figure Legend Snippet: Representative slice and streptavidin-Alexa labeled cells A . Photomicrograph of a slice from a Tg2576 mouse stained with thioflavin-S showing a low density of fibrillar plaques (arrowheads) in the cortex and hippocampus (inset: boxed area at higher magnification). B . Representative streptavidin-Alexa 488 labeled layer 3 pyramidal cells from Tg2576 (top) and Wt (bottom) mice. Scale bars = A: 1 mm, inset: 25 µm; B: 100 µm.

Techniques Used: Labeling, Staining, Mouse Assay

20) Product Images from "The Response of Cerebral Cortex to Haemorrhagic Damage: Experimental Evidence from a Penetrating Injury Model"

Article Title: The Response of Cerebral Cortex to Haemorrhagic Damage: Experimental Evidence from a Penetrating Injury Model

Journal: PLoS ONE

doi: 10.1371/journal.pone.0059740

Co-localisation of haem and amyloid labelling in human neocortex. A,B: The dark material is haem, identified with the Perls (Prussian blue) reaction, with DAB as the reporter molecule. The red labelling is Congo red, which labels amyloid. In the right hand panel the background Congo red labelling has been reduced. The largest haem deposit is surrounded by a shell of Congo red. Smaller haem deposits co-localise side-by-side with Congo red. C: Side-by-side co-localislation of haem and Congo red, in another plaque. D: Co-localisation of haem and Thioflavin S (green) in several plaques.
Figure Legend Snippet: Co-localisation of haem and amyloid labelling in human neocortex. A,B: The dark material is haem, identified with the Perls (Prussian blue) reaction, with DAB as the reporter molecule. The red labelling is Congo red, which labels amyloid. In the right hand panel the background Congo red labelling has been reduced. The largest haem deposit is surrounded by a shell of Congo red. Smaller haem deposits co-localise side-by-side with Congo red. C: Side-by-side co-localislation of haem and Congo red, in another plaque. D: Co-localisation of haem and Thioflavin S (green) in several plaques.

Techniques Used:

21) Product Images from "Glia Maturation Factor Expression in Hippocampus of Human Alzheimer's Disease"

Article Title: Glia Maturation Factor Expression in Hippocampus of Human Alzheimer's Disease

Journal: Neurochemical research

doi: 10.1007/s11064-013-1059-3

GMF IHC in the CA2 region of the hippocampus of AD brain. Hippocampal frozen sections from CA2 regions were immunostained with GMF followed by thioflavin-S histochemistry. CA2 hippocampal region show and GMF (A,B) and many APs (C,D arrows) immunoreactive
Figure Legend Snippet: GMF IHC in the CA2 region of the hippocampus of AD brain. Hippocampal frozen sections from CA2 regions were immunostained with GMF followed by thioflavin-S histochemistry. CA2 hippocampal region show and GMF (A,B) and many APs (C,D arrows) immunoreactive

Techniques Used: Immunohistochemistry

Up-regulated expression of GMF levels in the hippocampus of the AD brain. Hippocampal regions from AD brains were immunostained with antibody to GMF (A,B) followed by thioflavin-S histochemistry (C,D). The GMF expression levels (A,B) were increased in
Figure Legend Snippet: Up-regulated expression of GMF levels in the hippocampus of the AD brain. Hippocampal regions from AD brains were immunostained with antibody to GMF (A,B) followed by thioflavin-S histochemistry (C,D). The GMF expression levels (A,B) were increased in

Techniques Used: Expressing

22) Product Images from "Thioflavin S (NSC71948) Interferes with Bcl-2-Associated Athanogene (BAG-1)-Mediated Protein-Protein Interactions"

Article Title: Thioflavin S (NSC71948) Interferes with Bcl-2-Associated Athanogene (BAG-1)-Mediated Protein-Protein Interactions

Journal: The Journal of Pharmacology and Experimental Therapeutics

doi: 10.1124/jpet.109.153601

Identification of NSC71948 as an inhibitor of the in vitro BAG-1:HSC70 interaction. A, structure of NSC71948 (thioflavin S). B, inhibition of the in vitro BAG-1:HSC70 interaction by NSC71948. The experiment shows the mean of duplicate determinations (± S.D.) of BAG-1:HSC70 interaction in the presence of the indicated concentrations of NSC71948 (refilled stock). Experiment is representative of two independent experiments. C, luciferase counterscreen. The activity of recombinant luciferase was measured in the presence of NSC71948 or NSC119913 (both at 30 μM). The activity of luciferase in the absence of added compound was set at 100%. Data shown are mean (± S.D.) luciferase activity derived from two separate experiments, each performed in duplicate. D, BSA counterscreen. The ability of NSC71948 (30 μM) or NSC73413 (15 μM) to inhibit in vitro BAG-1:HSC70 binding was measured in the presence or absence of excess (0.1 mg/ml) BSA. Data shown are remaining activity (mean ± S.D.) in the presence of BSA and are derived from two separate experiments, each performed in duplicate.
Figure Legend Snippet: Identification of NSC71948 as an inhibitor of the in vitro BAG-1:HSC70 interaction. A, structure of NSC71948 (thioflavin S). B, inhibition of the in vitro BAG-1:HSC70 interaction by NSC71948. The experiment shows the mean of duplicate determinations (± S.D.) of BAG-1:HSC70 interaction in the presence of the indicated concentrations of NSC71948 (refilled stock). Experiment is representative of two independent experiments. C, luciferase counterscreen. The activity of recombinant luciferase was measured in the presence of NSC71948 or NSC119913 (both at 30 μM). The activity of luciferase in the absence of added compound was set at 100%. Data shown are mean (± S.D.) luciferase activity derived from two separate experiments, each performed in duplicate. D, BSA counterscreen. The ability of NSC71948 (30 μM) or NSC73413 (15 μM) to inhibit in vitro BAG-1:HSC70 binding was measured in the presence or absence of excess (0.1 mg/ml) BSA. Data shown are remaining activity (mean ± S.D.) in the presence of BSA and are derived from two separate experiments, each performed in duplicate.

Techniques Used: In Vitro, Inhibition, Luciferase, Activity Assay, Recombinant, Derivative Assay, Binding Assay

Analysis of structurally related compounds. (A) Structures of thioflavin S/NSC71948, thioflavin T and BTA1. (B) Inhibition of the in vitro BAG-1:HSC70 interaction. The experiment shows the mean (± S.D.) of duplicate determinations of BAG-1:HSC70 binding in the presence of the indicated concentrations of NSC71948 (♦), thioflavin S (×), thioflavin T (♢), BTA-1 (■), or DMSO (□). Binding in the absence of any added compound was set to 100%. The data are representative of two independent experiments. C, MCF7 cells were treated for 2 h with NSC71948, thioflavin S, thioflavin T, or BTA-1 (each at 50 μM), or DMSO as control. Expression of total and phosphorylated ERK1/2 was analyzed by immunoblotting. Data shown are representative of two experiments, each performed in duplicate. D, quantitation of effects of compounds (50 μM) on ERK1/2 phosphorylation. E, ZR-75-11 cells were treated with the indicated concentrations of NSC71948 (♦), thioflavin T (♢), BTA1 (■), or DMSO (□) as a control. After 6 days, cell number was determined by use of the CellTiter assay. Values obtained for untreated cells were set to 100%. Data are mean of triplicate determinations (± S.D.).
Figure Legend Snippet: Analysis of structurally related compounds. (A) Structures of thioflavin S/NSC71948, thioflavin T and BTA1. (B) Inhibition of the in vitro BAG-1:HSC70 interaction. The experiment shows the mean (± S.D.) of duplicate determinations of BAG-1:HSC70 binding in the presence of the indicated concentrations of NSC71948 (♦), thioflavin S (×), thioflavin T (♢), BTA-1 (■), or DMSO (□). Binding in the absence of any added compound was set to 100%. The data are representative of two independent experiments. C, MCF7 cells were treated for 2 h with NSC71948, thioflavin S, thioflavin T, or BTA-1 (each at 50 μM), or DMSO as control. Expression of total and phosphorylated ERK1/2 was analyzed by immunoblotting. Data shown are representative of two experiments, each performed in duplicate. D, quantitation of effects of compounds (50 μM) on ERK1/2 phosphorylation. E, ZR-75-11 cells were treated with the indicated concentrations of NSC71948 (♦), thioflavin T (♢), BTA1 (■), or DMSO (□) as a control. After 6 days, cell number was determined by use of the CellTiter assay. Values obtained for untreated cells were set to 100%. Data are mean of triplicate determinations (± S.D.).

Techniques Used: Inhibition, In Vitro, Binding Assay, Expressing, Quantitation Assay

23) Product Images from "Annexin A1 restores cerebrovascular integrity concomitant with reduced amyloid-β and tau pathology"

Article Title: Annexin A1 restores cerebrovascular integrity concomitant with reduced amyloid-β and tau pathology

Journal: Brain

doi: 10.1093/brain/awab050

Acute hrANXA1 treatment reduces cortical amyloid-β 40 pathology by increasing amyloid-β degradation in 5xFAD mice. ( A ) Left : ELISA analysis of amyloid-β 40 and amyloid-β 42 in the motor cortex (CTX) and hippocampus (HC) of 5xFAD mouse brain treated with hrANXA1 or vehicle, expressed as picograms per milligrams of protein ( n = 6–7/group). Right : Ratio of amyloid-β 42 and amyloid-β 40 detected by ELISA in the motor cortex and hippocampus. Unpaired two-tailed t -test. ( B ) Representative images and quantification of percentage area of Thioflavin-S staining in the cortex (CTX) and hippocampus (HC) of 5xFAD mice treated with hrANXA1 or vehicle ( n = 10–15 mice/group, mean of four to six sections analysed per mouse). ( C ) Scatter plot showing significant positive correlation between Evans blue dye content and Thioflavin-S staining percentage area in the hippocampus of 5xFAD mice acutely treated with 0.67 µg/kg hrANXA1 or vehicle at 3 months of age ( n = 7–8/group, linear regression analysis, Pearson’s r = 0.6337, P = 0.0112 (vehicle and ANXA1-treated). ( D ) Representative images and quantification of percentage area of anti-amyloid-β (6C3 antibody) in the cortex and hippocampus of 5xFAD mice treated with hrANXA1 or vehicle ( n = 8–11 mice/group, mean of four to six sections analysed per mouse). ( E ) Scatter plot showing significant positive correlation between Evans blue dye content and 6C3 percentage area stained in the hippocampus of 5xFAD mice acutely treated with 0.67 µg/kg hrANXA1 or vehicle at 3 months of age ( n = 5–6/group, linear regression analysis, Pearson’s r = 0.5401, P = 0.086). ( F ) Representative western blots and quantitative analysis of β-secretase expression and β-CTFs levels in 5xFAD mice treated with hrANXA1 or vehicle. BACE1 expression in the motor cortex, normalized to β-actin ( n = 9–10/group). β-CTF expression in the motor cortex, normalized to flAPP ( n = 10/group). ( G ) Representative western blots and quantitative analysis of neprilysin ( n = 6–7/group) and IDE expression ( n = 6/group) in the motor cortex of 5xFAD mice treated with hrANXA1 or vehicle, normalized to GAPDH. Unpaired two-tailed t -test. Columns represent mean ± SEM. * P
Figure Legend Snippet: Acute hrANXA1 treatment reduces cortical amyloid-β 40 pathology by increasing amyloid-β degradation in 5xFAD mice. ( A ) Left : ELISA analysis of amyloid-β 40 and amyloid-β 42 in the motor cortex (CTX) and hippocampus (HC) of 5xFAD mouse brain treated with hrANXA1 or vehicle, expressed as picograms per milligrams of protein ( n = 6–7/group). Right : Ratio of amyloid-β 42 and amyloid-β 40 detected by ELISA in the motor cortex and hippocampus. Unpaired two-tailed t -test. ( B ) Representative images and quantification of percentage area of Thioflavin-S staining in the cortex (CTX) and hippocampus (HC) of 5xFAD mice treated with hrANXA1 or vehicle ( n = 10–15 mice/group, mean of four to six sections analysed per mouse). ( C ) Scatter plot showing significant positive correlation between Evans blue dye content and Thioflavin-S staining percentage area in the hippocampus of 5xFAD mice acutely treated with 0.67 µg/kg hrANXA1 or vehicle at 3 months of age ( n = 7–8/group, linear regression analysis, Pearson’s r = 0.6337, P = 0.0112 (vehicle and ANXA1-treated). ( D ) Representative images and quantification of percentage area of anti-amyloid-β (6C3 antibody) in the cortex and hippocampus of 5xFAD mice treated with hrANXA1 or vehicle ( n = 8–11 mice/group, mean of four to six sections analysed per mouse). ( E ) Scatter plot showing significant positive correlation between Evans blue dye content and 6C3 percentage area stained in the hippocampus of 5xFAD mice acutely treated with 0.67 µg/kg hrANXA1 or vehicle at 3 months of age ( n = 5–6/group, linear regression analysis, Pearson’s r = 0.5401, P = 0.086). ( F ) Representative western blots and quantitative analysis of β-secretase expression and β-CTFs levels in 5xFAD mice treated with hrANXA1 or vehicle. BACE1 expression in the motor cortex, normalized to β-actin ( n = 9–10/group). β-CTF expression in the motor cortex, normalized to flAPP ( n = 10/group). ( G ) Representative western blots and quantitative analysis of neprilysin ( n = 6–7/group) and IDE expression ( n = 6/group) in the motor cortex of 5xFAD mice treated with hrANXA1 or vehicle, normalized to GAPDH. Unpaired two-tailed t -test. Columns represent mean ± SEM. * P

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Staining, Western Blot, Expressing

24) Product Images from "Retinal capillary degeneration and blood-retinal barrier disruption in murine models of Alzheimer’s disease"

Article Title: Retinal capillary degeneration and blood-retinal barrier disruption in murine models of Alzheimer’s disease

Journal: Acta Neuropathologica Communications

doi: 10.1186/s40478-020-01076-4

Retinal amyloidosis in blood vessel walls of APP SWE /PS1 ΔE9 (ADtg) mice. a–b Representative fluorescence images acquired using a 20 × or b 63 × microscope objectives, of isolated retinal microvasculature stained for Aβ (4G8, magenta), blood vessels (lectin, green), and nuclei (DAPI, blue) in age- and sex-matched perfused ADtg mice (n = 6) and wild-type littermates (WT; n = 6). Scale bars = 20 µm. c–d Representative fluorescence virtual cross-section images acquired using the Leica confocal microscope 63 × objective of the isolated retinal microvasculature stained for Aβ (4G8, magenta), blood vessels (lectin, green) and nuclei (DAPI, blue) in an 8-month-old male ADtg mouse. Scale bar = 20 µm. Arrows indicate vascular Aβ. e Quantitative analysis of Aβ (4G8)-immunoreactive (IR) area in each microscopic field of isolated retinal microvasculature from WT vs. ADtg mice. f Quantitative analysis of the Aβ (4G8)-IR area stratified by mice age group (8 months vs. 12 months) and genotype (WT vs. ADtg) in the same cohort. g–g′ . Representative fluorescence images of the isolated retinal microvasculature stained for Aβ (4G8, red), blood vessels (lectin, green) and nuclei (DAPI, blue) in an 8-month-old male ADtg mouse; Aβ signals occur in retinal vessel walls and vascular cells. h–i . Representative fluorescence images of fixed retinal cross-section stained for thioflavin-S (Thio-S, green), Aβ 40 (11A50-B10, red) and blood vessels (CD31, blue) in an 8-month-old male ADtg mouse showing h vertical blood vessel and i longitudinal blood vessel. Data from individual mice (circles) as well as group means ± SEMs are shown. Black-filled circles represent males and clear circles represent females. Fold changes are shown in red. * p
Figure Legend Snippet: Retinal amyloidosis in blood vessel walls of APP SWE /PS1 ΔE9 (ADtg) mice. a–b Representative fluorescence images acquired using a 20 × or b 63 × microscope objectives, of isolated retinal microvasculature stained for Aβ (4G8, magenta), blood vessels (lectin, green), and nuclei (DAPI, blue) in age- and sex-matched perfused ADtg mice (n = 6) and wild-type littermates (WT; n = 6). Scale bars = 20 µm. c–d Representative fluorescence virtual cross-section images acquired using the Leica confocal microscope 63 × objective of the isolated retinal microvasculature stained for Aβ (4G8, magenta), blood vessels (lectin, green) and nuclei (DAPI, blue) in an 8-month-old male ADtg mouse. Scale bar = 20 µm. Arrows indicate vascular Aβ. e Quantitative analysis of Aβ (4G8)-immunoreactive (IR) area in each microscopic field of isolated retinal microvasculature from WT vs. ADtg mice. f Quantitative analysis of the Aβ (4G8)-IR area stratified by mice age group (8 months vs. 12 months) and genotype (WT vs. ADtg) in the same cohort. g–g′ . Representative fluorescence images of the isolated retinal microvasculature stained for Aβ (4G8, red), blood vessels (lectin, green) and nuclei (DAPI, blue) in an 8-month-old male ADtg mouse; Aβ signals occur in retinal vessel walls and vascular cells. h–i . Representative fluorescence images of fixed retinal cross-section stained for thioflavin-S (Thio-S, green), Aβ 40 (11A50-B10, red) and blood vessels (CD31, blue) in an 8-month-old male ADtg mouse showing h vertical blood vessel and i longitudinal blood vessel. Data from individual mice (circles) as well as group means ± SEMs are shown. Black-filled circles represent males and clear circles represent females. Fold changes are shown in red. * p

Techniques Used: Mouse Assay, Fluorescence, Microscopy, Isolation, Staining

25) Product Images from "Inhibition of PKCδ reduces amyloid-β levels and reverses Alzheimer disease phenotypes"

Article Title: Inhibition of PKCδ reduces amyloid-β levels and reverses Alzheimer disease phenotypes

Journal: The Journal of Experimental Medicine

doi: 10.1084/jem.20171193

Rottlerin treatment improves cognitive deficits and reduces diffuse and fibrillar plaque burden and Aβ levels in APPswe/PS1dE9 mice. (A) Hidden-platform tests performed over 5 d in the Morris water maze. Significance was determined by repeated-measures ANOVA; vehicle-treated APPswe/PS1dE9 mice exhibited a longer escape latency compared with rottlerin- and vehicle-treated WT mice, whereas rottlerin-treated APPswe/PS1dE9 mice exhibited a shorter escape latency compared with vehicle-treated APPswe/PS1dE9 mice. (B) Probe trial 24 h after the last hidden-platform test in the Morris water maze. Percentage of time spent in the target quadrant is shown for APPswe/PS1dE9 and WT groups treated with rottlerin or vehicle. (C) Representative histological images from sagittal brain sections from APPswe/PS1dE9 mice treated with vehicle (top left) and rottlerin (top right) immunostained with monoclonal anti-Aβ antibody (DE2) for diffuse plaques (brown) and treated with vehicle (bottom left) and rottlerin (bottom right) stained with thioflavin S for fibrillar plaques (green). Bar, 300 µm. (D and E) Quantification of the percent area occupied by diffuse plaques (D) and fibrillar plaques (E) in APPswe/PS1dE9 mouse brain treated with vehicle and rottlerin. (F–H) Plaques from rottlerin-treated APPswe/PS1dE9 mice were found to be significantly lower than the vehicle-treated transgenic control group. Levels of soluble Aβ 1–40 (F), Aβ 1–42 (G), and Aβ oligomers (H) obtained in RIPA buffer in rottlerin-treated animals are significantly lower than in vehicle-treated controls. (I and J) Levels of insoluble Aβ 1–40 (I) and Aβ 1–42 (J) obtained by using formic acid extraction in rottlerin-treated animals are significantly lower than in vehicle-treated controls. n = 10 in each group. *, P
Figure Legend Snippet: Rottlerin treatment improves cognitive deficits and reduces diffuse and fibrillar plaque burden and Aβ levels in APPswe/PS1dE9 mice. (A) Hidden-platform tests performed over 5 d in the Morris water maze. Significance was determined by repeated-measures ANOVA; vehicle-treated APPswe/PS1dE9 mice exhibited a longer escape latency compared with rottlerin- and vehicle-treated WT mice, whereas rottlerin-treated APPswe/PS1dE9 mice exhibited a shorter escape latency compared with vehicle-treated APPswe/PS1dE9 mice. (B) Probe trial 24 h after the last hidden-platform test in the Morris water maze. Percentage of time spent in the target quadrant is shown for APPswe/PS1dE9 and WT groups treated with rottlerin or vehicle. (C) Representative histological images from sagittal brain sections from APPswe/PS1dE9 mice treated with vehicle (top left) and rottlerin (top right) immunostained with monoclonal anti-Aβ antibody (DE2) for diffuse plaques (brown) and treated with vehicle (bottom left) and rottlerin (bottom right) stained with thioflavin S for fibrillar plaques (green). Bar, 300 µm. (D and E) Quantification of the percent area occupied by diffuse plaques (D) and fibrillar plaques (E) in APPswe/PS1dE9 mouse brain treated with vehicle and rottlerin. (F–H) Plaques from rottlerin-treated APPswe/PS1dE9 mice were found to be significantly lower than the vehicle-treated transgenic control group. Levels of soluble Aβ 1–40 (F), Aβ 1–42 (G), and Aβ oligomers (H) obtained in RIPA buffer in rottlerin-treated animals are significantly lower than in vehicle-treated controls. (I and J) Levels of insoluble Aβ 1–40 (I) and Aβ 1–42 (J) obtained by using formic acid extraction in rottlerin-treated animals are significantly lower than in vehicle-treated controls. n = 10 in each group. *, P

Techniques Used: Mouse Assay, Staining, Transgenic Assay

26) Product Images from "The Disordered C-Terminus of Yeast Hsf1 Contains a Cryptic Low-Complexity Amyloidogenic Region"

Article Title: The Disordered C-Terminus of Yeast Hsf1 Contains a Cryptic Low-Complexity Amyloidogenic Region

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms19051384

Binding of amyloid dyes to the cryptic amyloid core of Hsf1. ( A ) Congo red (CR) spectral changes in the presence of Hsf1 peptide incubated at various concentrations during 120 h at 25 °C; note the characteristic shift from 497 nm to 515 nm when the dye is bound to amyloid-like aggregates; ( B ) changes in the fluorescence emission spectrum of Thioflavin-T (Th-T) when excited at 440 nm upon binding to the aggregated peptide at various concentrations after 120 h incubation at 25 °C; ( C ) Thioflavin-S fluorescence of stained aggregated amyloid material of Hsf1 at 100 μM in Phosphate buffered saline (PBS) buffer after 120 h incubation at 25 °C. Images were obtained at 40× magnification by phase contrast and fluorescence microscopy displaying the green fluorescence characteristic of amyloid structures.
Figure Legend Snippet: Binding of amyloid dyes to the cryptic amyloid core of Hsf1. ( A ) Congo red (CR) spectral changes in the presence of Hsf1 peptide incubated at various concentrations during 120 h at 25 °C; note the characteristic shift from 497 nm to 515 nm when the dye is bound to amyloid-like aggregates; ( B ) changes in the fluorescence emission spectrum of Thioflavin-T (Th-T) when excited at 440 nm upon binding to the aggregated peptide at various concentrations after 120 h incubation at 25 °C; ( C ) Thioflavin-S fluorescence of stained aggregated amyloid material of Hsf1 at 100 μM in Phosphate buffered saline (PBS) buffer after 120 h incubation at 25 °C. Images were obtained at 40× magnification by phase contrast and fluorescence microscopy displaying the green fluorescence characteristic of amyloid structures.

Techniques Used: Binding Assay, Incubation, Fluorescence, Staining, Microscopy

27) Product Images from "Assessment of brain beta-amyloid deposition in transgenic mouse models of Alzheimer’s disease with PET imaging agents 18F-flutemetamol and 18F-florbetaben"

Article Title: Assessment of brain beta-amyloid deposition in transgenic mouse models of Alzheimer’s disease with PET imaging agents 18F-flutemetamol and 18F-florbetaben

Journal: BMC Neuroscience

doi: 10.1186/s12868-018-0447-7

Visual overview of the thioflavin S staining images of a wild type mouse and b AD transgenic mouse. Left column shows DAPI (blue channel), middle column shows thioflavin S (green channel with specific staining signal) and right column shows merged image. Aβ deposits were found broadly in various brain regions including the cortex, hippocampus and thalamus in AD mice
Figure Legend Snippet: Visual overview of the thioflavin S staining images of a wild type mouse and b AD transgenic mouse. Left column shows DAPI (blue channel), middle column shows thioflavin S (green channel with specific staining signal) and right column shows merged image. Aβ deposits were found broadly in various brain regions including the cortex, hippocampus and thalamus in AD mice

Techniques Used: Staining, Transgenic Assay, Mouse Assay

Correlation with neuropathologic finding and visual PET images. a Paxinos and Franklinis the Mouse Brain in stereotaxic coordinates atlas representing our pathological section, b Thioflavin S staining image, c 18 F-florbetaben image, d 18 F-flutemetamol image. The 18 F-florbetaben PET images matched well to the thioflavin S staining image in aspects of signal intensity and spatial distribution pattern in cortical brain regions, compared with 18 F-flutemetamol images
Figure Legend Snippet: Correlation with neuropathologic finding and visual PET images. a Paxinos and Franklinis the Mouse Brain in stereotaxic coordinates atlas representing our pathological section, b Thioflavin S staining image, c 18 F-florbetaben image, d 18 F-flutemetamol image. The 18 F-florbetaben PET images matched well to the thioflavin S staining image in aspects of signal intensity and spatial distribution pattern in cortical brain regions, compared with 18 F-flutemetamol images

Techniques Used: Positron Emission Tomography, Staining

28) Product Images from "Granulocyte-macrophage colony-stimulating factor neuroprotective activities in Alzheimer’s disease mice"

Article Title: Granulocyte-macrophage colony-stimulating factor neuroprotective activities in Alzheimer’s disease mice

Journal: Journal of neuroimmunology

doi: 10.1016/j.jneuroim.2018.03.009

GM-CSF facilitates accumulation of microglia surrounding Aβ plaques in AD mice. ( A ) Representative images of Iba1 staining in the cortex of PBS- or GM-CSF-treated AD mice. Scale bar = 100μm. ( B ) Quantification of the mean number of Iba1-positive cells adjacent to thioflavin S (TS)-stained Aβ plaques (n=7 per group, 12 sections per brain, and 3 plaques per section). Data represent mean microglia number ± SEM, and ** p
Figure Legend Snippet: GM-CSF facilitates accumulation of microglia surrounding Aβ plaques in AD mice. ( A ) Representative images of Iba1 staining in the cortex of PBS- or GM-CSF-treated AD mice. Scale bar = 100μm. ( B ) Quantification of the mean number of Iba1-positive cells adjacent to thioflavin S (TS)-stained Aβ plaques (n=7 per group, 12 sections per brain, and 3 plaques per section). Data represent mean microglia number ± SEM, and ** p

Techniques Used: Mouse Assay, Staining

29) Product Images from "Holdase activity of secreted Hsp70 masks amyloid-β42 neurotoxicity in Drosophila"

Article Title: Holdase activity of secreted Hsp70 masks amyloid-β42 neurotoxicity in Drosophila

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.1608045113

Secreted Hsp70 suppresses Aβ42 neurotoxicity without Aβ42 clearance or disaggregation. ( A – H ) Accumulation of the amyloid dye thioflavin-S in the Drosophila brain in flies expressing LacZ or Aβ42 transgenes under the control
Figure Legend Snippet: Secreted Hsp70 suppresses Aβ42 neurotoxicity without Aβ42 clearance or disaggregation. ( A – H ) Accumulation of the amyloid dye thioflavin-S in the Drosophila brain in flies expressing LacZ or Aβ42 transgenes under the control

Techniques Used: Expressing

30) Product Images from "The APOE ɛ4/ɛ4 genotype potentiates vascular fibrin(ogen) deposition in amyloid-laden vessels in the brains of Alzheimer's disease patients"

Article Title: The APOE ɛ4/ɛ4 genotype potentiates vascular fibrin(ogen) deposition in amyloid-laden vessels in the brains of Alzheimer's disease patients

Journal: Journal of Cerebral Blood Flow & Metabolism

doi: 10.1038/jcbfm.2013.76

Alzheimer's disease (AD) apolipoprotein E ( APOE )  ɛ 4/ ɛ 4 cases present with significantly more fibrin(ogen) and cerebral amyloid angiopathy (CAA) double-positive vessels. Immunostaining of fibrin(ogen) and CAA was performed on cortical sections from AD brains. ( A ) Representative images of CAA (Thioflavin S staining, ThioS) (a, c, e) and fibrin(ogen) (b, d, f) in AD  APOE ɛ 2/ ɛ 3 (a, b), AD  APOE ɛ 3/ ɛ 3 (c, d), and AD  APOE ɛ 4/ ɛ 4 (e, f). Scale bar, 100  μ m. ( B ) Significantly more fibrin(ogen) and CAA double-positive vessels were observed in AD  APOE ɛ 4/ ɛ 4 cases compared with AD  APOE ɛ 3/ ɛ 3 ( P =0.02) or  ɛ 2/ ɛ 3 ( P =0.03) cases. Differences between groups were evaluated by one-way analysis of variance and Tukey's  post hoc  test. Values are presented as mean±s.e.m.
Figure Legend Snippet: Alzheimer's disease (AD) apolipoprotein E ( APOE ) ɛ 4/ ɛ 4 cases present with significantly more fibrin(ogen) and cerebral amyloid angiopathy (CAA) double-positive vessels. Immunostaining of fibrin(ogen) and CAA was performed on cortical sections from AD brains. ( A ) Representative images of CAA (Thioflavin S staining, ThioS) (a, c, e) and fibrin(ogen) (b, d, f) in AD APOE ɛ 2/ ɛ 3 (a, b), AD APOE ɛ 3/ ɛ 3 (c, d), and AD APOE ɛ 4/ ɛ 4 (e, f). Scale bar, 100  μ m. ( B ) Significantly more fibrin(ogen) and CAA double-positive vessels were observed in AD APOE ɛ 4/ ɛ 4 cases compared with AD APOE ɛ 3/ ɛ 3 ( P =0.02) or ɛ 2/ ɛ 3 ( P =0.03) cases. Differences between groups were evaluated by one-way analysis of variance and Tukey's post hoc test. Values are presented as mean±s.e.m.

Techniques Used: Cellular Antioxidant Activity Assay, Immunostaining, Staining

31) Product Images from "Hemorrhage promotes inflammation and myocardial damage following acute myocardial infarction: insights from a novel preclinical model and cardiovascular magnetic resonance"

Article Title: Hemorrhage promotes inflammation and myocardial damage following acute myocardial infarction: insights from a novel preclinical model and cardiovascular magnetic resonance

Journal: Journal of Cardiovascular Magnetic Resonance

doi: 10.1186/s12968-017-0361-7

Injury Patterns on Histology. Images show TTC, Thioflavin S and H E stained short axis sections obtained from explanted hearts at 24 h post intervention. TTC sections indicate region on necrosis appearing white in the I-HEM group (no hemorrhage) and reddish white in I+HEM group (hemorrhage); the +HEM group did no show infarction. Under ultra-violet light, non-fluorescent regions on the Thioflavin S stain highlight areas of compromised endothelium as seen in the +HEM and I+HEM groups; this confirms presence of microvascular damage in the hemorrhage groups. The H E images were obtained from the region of interest shown on the TTC stained sections. Hemorrhage was apparent in the +HEM and I+HEM groups as evidenced from the interstitial distribution of red blood cells ( open arrow heads ) in the affected LAD territory; red blood cells were absent in the I-HEM group. Edematous development was observed in all three groups ( arrows ) along with the presence of inflammatory cells ( closed arrow heads )
Figure Legend Snippet: Injury Patterns on Histology. Images show TTC, Thioflavin S and H E stained short axis sections obtained from explanted hearts at 24 h post intervention. TTC sections indicate region on necrosis appearing white in the I-HEM group (no hemorrhage) and reddish white in I+HEM group (hemorrhage); the +HEM group did no show infarction. Under ultra-violet light, non-fluorescent regions on the Thioflavin S stain highlight areas of compromised endothelium as seen in the +HEM and I+HEM groups; this confirms presence of microvascular damage in the hemorrhage groups. The H E images were obtained from the region of interest shown on the TTC stained sections. Hemorrhage was apparent in the +HEM and I+HEM groups as evidenced from the interstitial distribution of red blood cells ( open arrow heads ) in the affected LAD territory; red blood cells were absent in the I-HEM group. Edematous development was observed in all three groups ( arrows ) along with the presence of inflammatory cells ( closed arrow heads )

Techniques Used: Staining

32) Product Images from "Different curcumin forms selectively bind fibrillar amyloid beta in post mortem Alzheimer’s disease brains: Implications for in-vivo diagnostics"

Article Title: Different curcumin forms selectively bind fibrillar amyloid beta in post mortem Alzheimer’s disease brains: Implications for in-vivo diagnostics

Journal: Acta Neuropathologica Communications

doi: 10.1186/s40478-018-0577-2

Staining of different curcumin forms to amyloid plaques, neurofibrillary tangles and CAA. Staining of amyloid-beta plaques, neurofibrillary tangles and CAA with Thioflavin-S and curcuminoids ( a ), conjugates ( b ) and bio-available forms ( c ). Scale bars 100 μm
Figure Legend Snippet: Staining of different curcumin forms to amyloid plaques, neurofibrillary tangles and CAA. Staining of amyloid-beta plaques, neurofibrillary tangles and CAA with Thioflavin-S and curcuminoids ( a ), conjugates ( b ) and bio-available forms ( c ). Scale bars 100 μm

Techniques Used: Staining, Cellular Antioxidant Activity Assay

33) Product Images from "Induction of the unfolded protein response and cell death pathway in Alzheimer's disease, but not in aged Tg2576 mice"

Article Title: Induction of the unfolded protein response and cell death pathway in Alzheimer's disease, but not in aged Tg2576 mice

Journal: Experimental & Molecular Medicine

doi: 10.3858/emm.2010.42.5.040

Induction of the unfolded protein response by ER stress in AD. (A, B) RT-PCR analysis of XBP-1 mRNA splicing in the temporal cortex of control (CTRL) and AD brain (A). XBP-1 mRNA splicing was analyzed by scaling intensity of splicing band (193bp) to total band (193 bp + 219 bp) (B). (C) Representative images of thioflavin S staining in the temporal cortex of control (CTRL) and AD brain. (D-F) Western blot analysis of PDI, GRP78, and actin (D). Levels of PDI (E) and GRP78 (F) were measured and normalized to the level of relevant actin ( n  = 4 for control and  n  = 8 for AD).
Figure Legend Snippet: Induction of the unfolded protein response by ER stress in AD. (A, B) RT-PCR analysis of XBP-1 mRNA splicing in the temporal cortex of control (CTRL) and AD brain (A). XBP-1 mRNA splicing was analyzed by scaling intensity of splicing band (193bp) to total band (193 bp + 219 bp) (B). (C) Representative images of thioflavin S staining in the temporal cortex of control (CTRL) and AD brain. (D-F) Western blot analysis of PDI, GRP78, and actin (D). Levels of PDI (E) and GRP78 (F) were measured and normalized to the level of relevant actin ( n = 4 for control and n = 8 for AD).

Techniques Used: Reverse Transcription Polymerase Chain Reaction, Staining, Western Blot

Neither the UPR nor cell death pathway is induced in aged Tg2576 mice. (A) Fluorescence photomicrographs of cortical sections from wild type (WT) and Tg2576 mice at the age of 17 months after thioflavin-S staining. (B) Western blot analysis of PDI, GRP78, cleaved caspase-3, 9, 12, CHOP, and actin in the cortex of wild type and Tg2576 mice.
Figure Legend Snippet: Neither the UPR nor cell death pathway is induced in aged Tg2576 mice. (A) Fluorescence photomicrographs of cortical sections from wild type (WT) and Tg2576 mice at the age of 17 months after thioflavin-S staining. (B) Western blot analysis of PDI, GRP78, cleaved caspase-3, 9, 12, CHOP, and actin in the cortex of wild type and Tg2576 mice.

Techniques Used: Mouse Assay, Fluorescence, Staining, Western Blot

34) Product Images from "Cyclooxygenase-1 inhibition reduces amyloid pathology and improves memory deficits in a mouse model of Alzheimer's disease"

Article Title: Cyclooxygenase-1 inhibition reduces amyloid pathology and improves memory deficits in a mouse model of Alzheimer's disease

Journal: Journal of neurochemistry

doi: 10.1111/jnc.12059

SC-560 treatment reduces amyloid deposits. (a, b) Representative images of Congo red (a) and thioflavin S staining (b) in hippocampal subiculum in 3 × Tg-AD mice treated with vehicle or SC-560. Scale bar, 100 μm. Boxed regions (left) indicate
Figure Legend Snippet: SC-560 treatment reduces amyloid deposits. (a, b) Representative images of Congo red (a) and thioflavin S staining (b) in hippocampal subiculum in 3 × Tg-AD mice treated with vehicle or SC-560. Scale bar, 100 μm. Boxed regions (left) indicate

Techniques Used: Staining, Mouse Assay

35) Product Images from "Amyloid Properties of the Mouse Egg Zona Pellucida"

Article Title: Amyloid Properties of the Mouse Egg Zona Pellucida

Journal: PLoS ONE

doi: 10.1371/journal.pone.0129907

Amyloidogenic properties of the mouse ZP. A) The detection of amyloids in isolated mouse ZP was carried out using the amyloid conformation-dependent antibodies anti-fibrillar OC and anti-oligomer A11 in immunofluorescence analysis. Normal rabbit serum (NRS) served as a control. The anti-ZP3 antibody was used as a marker for the ZP with normal goat IgG serving as a control antibody. The corresponding phase images are shown for each fluorescent image. B) Intact ZP stained with 0.1% thioflavin S to detect amyloids. Scale bar = 50 μm. C) ZP pellets stained with 0.2% Congo Red showed yellow-green birefringence (arrow) when examined under polarizing light and bright red fluorescence when examined with UV light. Scale bar = 10 μm.
Figure Legend Snippet: Amyloidogenic properties of the mouse ZP. A) The detection of amyloids in isolated mouse ZP was carried out using the amyloid conformation-dependent antibodies anti-fibrillar OC and anti-oligomer A11 in immunofluorescence analysis. Normal rabbit serum (NRS) served as a control. The anti-ZP3 antibody was used as a marker for the ZP with normal goat IgG serving as a control antibody. The corresponding phase images are shown for each fluorescent image. B) Intact ZP stained with 0.1% thioflavin S to detect amyloids. Scale bar = 50 μm. C) ZP pellets stained with 0.2% Congo Red showed yellow-green birefringence (arrow) when examined under polarizing light and bright red fluorescence when examined with UV light. Scale bar = 10 μm.

Techniques Used: Isolation, Immunofluorescence, Marker, Staining, Fluorescence

36) Product Images from "A nontransgenic mouse model shows inducible amyloid-? (A?) peptide deposition and elucidates the role of apolipoprotein E in the amyloid cascade"

Article Title: A nontransgenic mouse model shows inducible amyloid-? (A?) peptide deposition and elucidates the role of apolipoprotein E in the amyloid cascade

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.0404458101

Quantitative determination of the numbers of Aβ42 ( A ), Aβ40 ( B ), and fibrillar Aβ ( C ) deposits in brains of apoE4-transgenic (ApoE4-tg), apoE3-transgenic (ApoE3-tg), apoE-deficient (ApoE-def.), and control mice after i.c.v. infusion of thiorphan for 1 month (see Materials and Methods ). Coronal sections at the level of maximal Aβ deposition (bregma –2.8) were stained with anti-Aβ42 (mAb G211) or anti-Aβ40 (mAb G210) antibodies or with thioflavin-S (see Materials and Methods ). Values (means ± SD of four or five mice per group) for each of the Aβ deposition measurements are for two sections per mouse. * , P
Figure Legend Snippet: Quantitative determination of the numbers of Aβ42 ( A ), Aβ40 ( B ), and fibrillar Aβ ( C ) deposits in brains of apoE4-transgenic (ApoE4-tg), apoE3-transgenic (ApoE3-tg), apoE-deficient (ApoE-def.), and control mice after i.c.v. infusion of thiorphan for 1 month (see Materials and Methods ). Coronal sections at the level of maximal Aβ deposition (bregma –2.8) were stained with anti-Aβ42 (mAb G211) or anti-Aβ40 (mAb G210) antibodies or with thioflavin-S (see Materials and Methods ). Values (means ± SD of four or five mice per group) for each of the Aβ deposition measurements are for two sections per mouse. * , P

Techniques Used: Transgenic Assay, Mouse Assay, Staining

Rostrocaudal distribution of Aβ42 ( A ), Aβ40 ( B ), and fibrillar Aβ ( C ) deposits in brains of apoE-transgenic and control mice after inhibition of neprilysin. ApoE4-transgenic (•), apoE3-transgenic (○), apoE-deficient (▵), and control (▴) mice were infused i.c.v. with thiorphan for 1 month (see Materials and Methods ). Brains were then cut coronally (–4 mm to +3 mm from bregma), and consecutive coronal sections at the indicated positions were stained for Aβ42 (mAb G211), Aβ40 (mAb G210), or fibrillar Aβ deposits (thioflavin-S) (see Materials and Methods ). Values (means ± SD of four or five mice per group) are for two sections per mouse at each of the indicated bregma levels.
Figure Legend Snippet: Rostrocaudal distribution of Aβ42 ( A ), Aβ40 ( B ), and fibrillar Aβ ( C ) deposits in brains of apoE-transgenic and control mice after inhibition of neprilysin. ApoE4-transgenic (•), apoE3-transgenic (○), apoE-deficient (▵), and control (▴) mice were infused i.c.v. with thiorphan for 1 month (see Materials and Methods ). Brains were then cut coronally (–4 mm to +3 mm from bregma), and consecutive coronal sections at the indicated positions were stained for Aβ42 (mAb G211), Aβ40 (mAb G210), or fibrillar Aβ deposits (thioflavin-S) (see Materials and Methods ). Values (means ± SD of four or five mice per group) are for two sections per mouse at each of the indicated bregma levels.

Techniques Used: Transgenic Assay, Mouse Assay, Inhibition, Staining

Effects of the apoE genotype on Aβ deposition after inhibition of neprilysin. ( A ) Representative micrographs of cortical fields of coronal brain sections (bregma –2.8 mm) from apoE4-(ApoE4-tg.) and apoE3-(ApoE3-tg.) transgenic, apoE-deficient (ApoE-def.), and control mice infused i.c.v. for 1 month with the neprilysin inhibitor thiorphan (see Materials and Methods ). ( B ) Representative immunoblots of soluble (SAβ) and insoluble (InAβ) Aβ of the same groups of three mice each. Immunohistochemistry (anti-Aβ42 mAb G211 and anti Aβ40 mAb G210), histochemistry (thioflavin-S), and immunoblot analysis (anti-Aβ mAb 4G8) were performed as described in Materials and Methods . (Scale bar, 50 μm.)
Figure Legend Snippet: Effects of the apoE genotype on Aβ deposition after inhibition of neprilysin. ( A ) Representative micrographs of cortical fields of coronal brain sections (bregma –2.8 mm) from apoE4-(ApoE4-tg.) and apoE3-(ApoE3-tg.) transgenic, apoE-deficient (ApoE-def.), and control mice infused i.c.v. for 1 month with the neprilysin inhibitor thiorphan (see Materials and Methods ). ( B ) Representative immunoblots of soluble (SAβ) and insoluble (InAβ) Aβ of the same groups of three mice each. Immunohistochemistry (anti-Aβ42 mAb G211 and anti Aβ40 mAb G210), histochemistry (thioflavin-S), and immunoblot analysis (anti-Aβ mAb 4G8) were performed as described in Materials and Methods . (Scale bar, 50 μm.)

Techniques Used: Inhibition, Transgenic Assay, Mouse Assay, Western Blot, Immunohistochemistry

37) Product Images from "Maysin and Its Flavonoid Derivative from Centipedegrass Attenuates Amyloid Plaques by Inducting Humoral Immune Response with Th2 Skewed Cytokine Response in the Tg (APPswe, PS1dE9) Alzheimer’s Mouse Model"

Article Title: Maysin and Its Flavonoid Derivative from Centipedegrass Attenuates Amyloid Plaques by Inducting Humoral Immune Response with Th2 Skewed Cytokine Response in the Tg (APPswe, PS1dE9) Alzheimer’s Mouse Model

Journal: PLoS ONE

doi: 10.1371/journal.pone.0169509

Comparison of the amyloid burden in the Tg brains by the CG extracts. (A) The amyloid plaques were identified by immunohistochemistry (IH) and H E counterstaining. The CG extract-treated (EA-CG fraction) and control Tg brain sections were formalin fixed and subjected to immunohistochemistry with the 6E10 antibody that interacts with the Aβ1–16 epitope (monoclonal antibody) in the hippocampus (a, control; b, EA-CG treated) and cortex (e, control; f, EA-CG-treated). Concomitant with the IH staining, other hippocampal (n = 6) (c, vehicle; d, EA-CG-treated) and cortical (n = 6) (g, vehicle; h, EA-CG-treated) sections from the Mo/Hu APPswe PS1dE9 mice were subjected to H E counterstaining. The scale bar indicates 200 μm (a through h). The amyloid load (% of stained area) as a quantitation result was decreased by the CG extracts (B. hippocampus area; C. cortex area). The area in the hippocampus (D, vehicle alone vs. EA-CG) and cortex (E, vehicle alone vs. EA-CG) was quantified by comparing the morphometric analysis of the Thioflavin-S-stained area between the vehicle-treated vs. EA-CG-treated Mo/Hu APPswe PS1dE9 brain sections. The values (%) shown were the means ± S.E.M. *p
Figure Legend Snippet: Comparison of the amyloid burden in the Tg brains by the CG extracts. (A) The amyloid plaques were identified by immunohistochemistry (IH) and H E counterstaining. The CG extract-treated (EA-CG fraction) and control Tg brain sections were formalin fixed and subjected to immunohistochemistry with the 6E10 antibody that interacts with the Aβ1–16 epitope (monoclonal antibody) in the hippocampus (a, control; b, EA-CG treated) and cortex (e, control; f, EA-CG-treated). Concomitant with the IH staining, other hippocampal (n = 6) (c, vehicle; d, EA-CG-treated) and cortical (n = 6) (g, vehicle; h, EA-CG-treated) sections from the Mo/Hu APPswe PS1dE9 mice were subjected to H E counterstaining. The scale bar indicates 200 μm (a through h). The amyloid load (% of stained area) as a quantitation result was decreased by the CG extracts (B. hippocampus area; C. cortex area). The area in the hippocampus (D, vehicle alone vs. EA-CG) and cortex (E, vehicle alone vs. EA-CG) was quantified by comparing the morphometric analysis of the Thioflavin-S-stained area between the vehicle-treated vs. EA-CG-treated Mo/Hu APPswe PS1dE9 brain sections. The values (%) shown were the means ± S.E.M. *p

Techniques Used: Immunohistochemistry, Staining, Mouse Assay, Quantitation Assay

38) Product Images from "Swarming motility and biofilm formation of Paenibacillus larvae, the etiological agent of American Foulbrood of honey bees (Apis mellifera)"

Article Title: Swarming motility and biofilm formation of Paenibacillus larvae, the etiological agent of American Foulbrood of honey bees (Apis mellifera)

Journal: Scientific Reports

doi: 10.1038/s41598-018-27193-8

Fluorescence microscopy of P. larvae floating biofilms. Bacterial suspensions of P. larvae ERIC I (ATCC9545; A , C ) and P. larvae ERIC II (DSM25430; B , D ) in Sf-900 II SFM medium supplemented with 30 µg/ml thioflavin S were incubated without agitation in 96-well-plates at 37 °C for six days. The thioflavin S-stained extracellular matrix in the floating biofilms was visualized using fluorescence microscopy; Z-stack processing was performed to obtain three-dimensional images of the wells containing the biofilms ( A , B ) and the region within the wells where the biofilms were located ( C , D ). Bars represent 20 µm.
Figure Legend Snippet: Fluorescence microscopy of P. larvae floating biofilms. Bacterial suspensions of P. larvae ERIC I (ATCC9545; A , C ) and P. larvae ERIC II (DSM25430; B , D ) in Sf-900 II SFM medium supplemented with 30 µg/ml thioflavin S were incubated without agitation in 96-well-plates at 37 °C for six days. The thioflavin S-stained extracellular matrix in the floating biofilms was visualized using fluorescence microscopy; Z-stack processing was performed to obtain three-dimensional images of the wells containing the biofilms ( A , B ) and the region within the wells where the biofilms were located ( C , D ). Bars represent 20 µm.

Techniques Used: Fluorescence, Microscopy, Incubation, Staining

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    Millipore thioflavin s
    Rac1 activity decreases α-SYN accumulation and aggregation in the neuroblastoma cell line BE(2)-M17. a Representative confocal images of α-SYN over-expressing cells induced with 10 μM retinoic acid (RA) and treated with 10 mM sodium butyrate (SB) for 36 h. Cells were transduced with Control-GFP (upper row), RAC1 (WT)-GFP (middle row) and RAC1 (CA)-GFP (bottom row), and co-stained for <t>Thioflavin</t> S (green) and α-SYN (red). Arrows indicate Thioflavin S positive aggregates with amyloidal structure. b Bar graph showing the quantitative analyses of the neuronal soma area (in percentage %) covered by Thioflavin S positive stain in individual cells transduced with (WT)- or (CA) RAC1 or with the corresponding control. N = 14 (EV), N = 25 (WT), and N = 24 (CA), from at least three independent experiments. Data are presented as mean ± SEM. Statistics, *** P
    Thioflavin S, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    IL-1Ra immunoreactivity is detectable in human islet α-cells. (A)  Paraffin-embedded sections from human islets transduced with Ad-prohIAPP-siRNA (or Ad-control-siRNA) and cultured (11.1 mmol/L glucose) for 7 days were immunolabeled for glucagon (red) and IL-1Ra (green).  (B)  Triple immunostaining of pre-culture human islets for insulin (blue), glucagon (red) and IL-1Ra (green). The squares (dashed white lines) denote regions enlarged and depicted as inserts at the bottom right of the corresponding images. Amyloid formation during islet culture is shown as an insert in merged micrographs (top right, insulin; red and thioflavin S; blue). Scale bar = 50 μm; inserts: ×3. Micrographs are representative of four independent studies (4 human islet preparations).
    Thioflavin S Solution, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/thioflavin s solution/product/Millipore
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    Rac1 activity decreases α-SYN accumulation and aggregation in the neuroblastoma cell line BE(2)-M17. a Representative confocal images of α-SYN over-expressing cells induced with 10 μM retinoic acid (RA) and treated with 10 mM sodium butyrate (SB) for 36 h. Cells were transduced with Control-GFP (upper row), RAC1 (WT)-GFP (middle row) and RAC1 (CA)-GFP (bottom row), and co-stained for Thioflavin S (green) and α-SYN (red). Arrows indicate Thioflavin S positive aggregates with amyloidal structure. b Bar graph showing the quantitative analyses of the neuronal soma area (in percentage %) covered by Thioflavin S positive stain in individual cells transduced with (WT)- or (CA) RAC1 or with the corresponding control. N = 14 (EV), N = 25 (WT), and N = 24 (CA), from at least three independent experiments. Data are presented as mean ± SEM. Statistics, *** P

    Journal: Molecular Neurobiology

    Article Title: The Small GTPase RAC1/CED-10 Is Essential in Maintaining Dopaminergic Neuron Function and Survival Against α-Synuclein-Induced Toxicity

    doi: 10.1007/s12035-018-0881-7

    Figure Lengend Snippet: Rac1 activity decreases α-SYN accumulation and aggregation in the neuroblastoma cell line BE(2)-M17. a Representative confocal images of α-SYN over-expressing cells induced with 10 μM retinoic acid (RA) and treated with 10 mM sodium butyrate (SB) for 36 h. Cells were transduced with Control-GFP (upper row), RAC1 (WT)-GFP (middle row) and RAC1 (CA)-GFP (bottom row), and co-stained for Thioflavin S (green) and α-SYN (red). Arrows indicate Thioflavin S positive aggregates with amyloidal structure. b Bar graph showing the quantitative analyses of the neuronal soma area (in percentage %) covered by Thioflavin S positive stain in individual cells transduced with (WT)- or (CA) RAC1 or with the corresponding control. N = 14 (EV), N = 25 (WT), and N = 24 (CA), from at least three independent experiments. Data are presented as mean ± SEM. Statistics, *** P

    Article Snippet: After incubation with the antibodies, coverslips were immersed in 0.005% thioflavin S (Sigma-Aldrich, Madrid, Spain) in PBS for 8 min and then rinsed twice in ethanol 70% and once in PBS.

    Techniques: Activity Assay, Expressing, Transduction, Staining

    Insoluble proteins form NFT-like aggregates rapidly after OA injection at both the injection site and distal brain regions. a β-pleated sheet protein structures are identified by thioflavin-S. Intracellular aggregates are observed at both the injection site and distal sites in the somatosensory cortex at 24 h, which persist 7 days post injection. Scale bar = 50 μM. b High magnification of thioflavin-S positive cells in the amygdala and somatosensory cortex. Scale bar = 10 μm. c Coronal sections taken from OA injected MAPT KO show no phospho-tau immunoreactivity. d No thioflavin positive structures are observed in MAPT KO mice with background staining comparable to the wild-type controls at 24 h. Scale bar = 50 μm

    Journal: Acta Neuropathologica Communications

    Article Title: A local insult of okadaic acid in wild-type mice induces tau phosphorylation and protein aggregation in anatomically distinct brain regions

    doi: 10.1186/s40478-016-0300-0

    Figure Lengend Snippet: Insoluble proteins form NFT-like aggregates rapidly after OA injection at both the injection site and distal brain regions. a β-pleated sheet protein structures are identified by thioflavin-S. Intracellular aggregates are observed at both the injection site and distal sites in the somatosensory cortex at 24 h, which persist 7 days post injection. Scale bar = 50 μM. b High magnification of thioflavin-S positive cells in the amygdala and somatosensory cortex. Scale bar = 10 μm. c Coronal sections taken from OA injected MAPT KO show no phospho-tau immunoreactivity. d No thioflavin positive structures are observed in MAPT KO mice with background staining comparable to the wild-type controls at 24 h. Scale bar = 50 μm

    Article Snippet: Slides were incubated in filtered 1 % thioflavin-S (Sigma) in 80 % ethanol for 15 min at room temperature (RT), protected from light.

    Techniques: Injection, Mouse Assay, Staining

    EFhd2 forms amyloid structures in vitro A , Various concentrations of purified, recombinant His EFhd2 WT (7.5, 30, 40μM) were incubated at 37°C with heparin and DTT (dash, dotted, solid line, respectively). After 20 hrs, samples were added Thioflavin-S (ThioS) and intensity was read in a fluorimeter at Exc.440 nm and Em.550 nm. ThioS stain has a high affinity for cross-β containing structures such as amyloids. EFhd2 showed affinity for this stain and an increase in ThioS binding was observed in a concentration-dependent manner. Baseline ThioS signal of buffer with heparin and DTT is shown (gray line) for comparison with EFhd2-containing reactions. B, Recombinant His EFhd2 WT (40μM) can also bind ThioS stain when incubated without heparin. Formation of cross-β structures by His EFhd2 WT was affected upon the addition of calcium, as it shows a reduction in the binding of ThioS (solid line, minus calcium; dashed lines, plus 1mM CaCl 2 ). Baseline ThioS signal of buffers with heparin and DTT minus (gray line) and plus (dotted line) 1mM CaCl 2 are shown for comparison with protein-containing reactions.

    Journal: Journal of neurochemistry

    Article Title: EFHD2 IS A NOVEL AMYLOID PROTEIN ASSOCIATED TO PATHOLOGICAL TAU IN ALZHEIMER'S DISEASE

    doi: 10.1111/jnc.12155

    Figure Lengend Snippet: EFhd2 forms amyloid structures in vitro A , Various concentrations of purified, recombinant His EFhd2 WT (7.5, 30, 40μM) were incubated at 37°C with heparin and DTT (dash, dotted, solid line, respectively). After 20 hrs, samples were added Thioflavin-S (ThioS) and intensity was read in a fluorimeter at Exc.440 nm and Em.550 nm. ThioS stain has a high affinity for cross-β containing structures such as amyloids. EFhd2 showed affinity for this stain and an increase in ThioS binding was observed in a concentration-dependent manner. Baseline ThioS signal of buffer with heparin and DTT is shown (gray line) for comparison with EFhd2-containing reactions. B, Recombinant His EFhd2 WT (40μM) can also bind ThioS stain when incubated without heparin. Formation of cross-β structures by His EFhd2 WT was affected upon the addition of calcium, as it shows a reduction in the binding of ThioS (solid line, minus calcium; dashed lines, plus 1mM CaCl 2 ). Baseline ThioS signal of buffers with heparin and DTT minus (gray line) and plus (dotted line) 1mM CaCl 2 are shown for comparison with protein-containing reactions.

    Article Snippet: After incubation, 0.5mM Thioflavin-S (SIGMA) was added to the samples and analyzed in a fluorimeter (Cary Eclipse fluorimeter, Agilent Technologies) with a temperature-controlled cell holder set at 37°C.

    Techniques: In Vitro, Purification, Recombinant, Incubation, Staining, Binding Assay, Concentration Assay

    IL-1Ra immunoreactivity is detectable in human islet α-cells. (A)  Paraffin-embedded sections from human islets transduced with Ad-prohIAPP-siRNA (or Ad-control-siRNA) and cultured (11.1 mmol/L glucose) for 7 days were immunolabeled for glucagon (red) and IL-1Ra (green).  (B)  Triple immunostaining of pre-culture human islets for insulin (blue), glucagon (red) and IL-1Ra (green). The squares (dashed white lines) denote regions enlarged and depicted as inserts at the bottom right of the corresponding images. Amyloid formation during islet culture is shown as an insert in merged micrographs (top right, insulin; red and thioflavin S; blue). Scale bar = 50 μm; inserts: ×3. Micrographs are representative of four independent studies (4 human islet preparations).

    Journal: Molecular Metabolism

    Article Title: Amyloid formation disrupts the balance between interleukin-1β and interleukin-1 receptor antagonist in human islets

    doi: 10.1016/j.molmet.2017.05.016

    Figure Lengend Snippet: IL-1Ra immunoreactivity is detectable in human islet α-cells. (A) Paraffin-embedded sections from human islets transduced with Ad-prohIAPP-siRNA (or Ad-control-siRNA) and cultured (11.1 mmol/L glucose) for 7 days were immunolabeled for glucagon (red) and IL-1Ra (green). (B) Triple immunostaining of pre-culture human islets for insulin (blue), glucagon (red) and IL-1Ra (green). The squares (dashed white lines) denote regions enlarged and depicted as inserts at the bottom right of the corresponding images. Amyloid formation during islet culture is shown as an insert in merged micrographs (top right, insulin; red and thioflavin S; blue). Scale bar = 50 μm; inserts: ×3. Micrographs are representative of four independent studies (4 human islet preparations).

    Article Snippet: For thioflavin S staining, sections were incubated with 0.5% (wt/vol.) thioflavin S solution (Sigma–Aldrich, Oakville, ON, Canada) for 5 min at room temperature.

    Techniques: Transduction, Cell Culture, Immunolabeling, Triple Immunostaining

    IL-1Ra levels are increased in oligomer-positive islet areas and reduced in thioflavin S-positive islet areas. (A) Paraffin embedded sections from 7-day cultured (11.1 mmol/L glucose) non-transduced or transduced with Ad-prohIAPP-siRNA (or Ad-control-siRNA) islets were immunolabeled for insulin (red) and IL-1Ra (green). The squares (dashed white lines) denote enlarged regions shown as inserts. (B) Human islets were immunolabeled for IL-1Ra (green) and A11 (red; top panel) or insulin (red), IL-1Ra (green) and thioflavin S (Thio S; blue; bottom panel). (C) Paraffin-embedded sections from (left to right): ob/ob mouse and human adipose tissue immunolabeled for IL-1Ra (green; positive control); human islets incubated with secondary antibody alone (negative control) and anakinra-treated human islets immunolabeled for insulin (red) and IL-1Ra (green). Scale bar = 50 μm; inserts: ×3 (A11: ×4). Micrographs are representative of four independent studies (4 human islet preparations).

    Journal: Molecular Metabolism

    Article Title: Amyloid formation disrupts the balance between interleukin-1β and interleukin-1 receptor antagonist in human islets

    doi: 10.1016/j.molmet.2017.05.016

    Figure Lengend Snippet: IL-1Ra levels are increased in oligomer-positive islet areas and reduced in thioflavin S-positive islet areas. (A) Paraffin embedded sections from 7-day cultured (11.1 mmol/L glucose) non-transduced or transduced with Ad-prohIAPP-siRNA (or Ad-control-siRNA) islets were immunolabeled for insulin (red) and IL-1Ra (green). The squares (dashed white lines) denote enlarged regions shown as inserts. (B) Human islets were immunolabeled for IL-1Ra (green) and A11 (red; top panel) or insulin (red), IL-1Ra (green) and thioflavin S (Thio S; blue; bottom panel). (C) Paraffin-embedded sections from (left to right): ob/ob mouse and human adipose tissue immunolabeled for IL-1Ra (green; positive control); human islets incubated with secondary antibody alone (negative control) and anakinra-treated human islets immunolabeled for insulin (red) and IL-1Ra (green). Scale bar = 50 μm; inserts: ×3 (A11: ×4). Micrographs are representative of four independent studies (4 human islet preparations).

    Article Snippet: For thioflavin S staining, sections were incubated with 0.5% (wt/vol.) thioflavin S solution (Sigma–Aldrich, Oakville, ON, Canada) for 5 min at room temperature.

    Techniques: Cell Culture, Transduction, Immunolabeling, Positive Control, Incubation, Negative Control

    Treatment with neutralizing IL-1β antibody markedly reduces islet IL-1β immunoreactivity and prevents β-cell Fas upregulation induced by amyloid formation in cultured human islets. Human islets non-transduced or transduced with Ad-prohIAPP-siRNA or Ad-control-siRNA (MOI: 20) were cultured with (+nIL1β) or without (-nIL1β) a neutralizing monoclonal human IL-1β antibody (1 μg/ml) in CMRL (11.1 mmol/L glucose) for 7 days. (A) Paraffin-embedded islet sections were immunolabeled for insulin (red), IL-1β or Fas (green) and thioflavin S (Thio S; blue) as indicated. The squares (dashed white lines) denote regions enlarged and depicted as inserts at the bottom right of each image. The top right inserts in each micrograph show immunolabeling for insulin (green) and A11 (red). Scale bar = 50 μm; inserts: ×3 (A11: ×4). (B) The proportion of Fas positive islet β-cells (fold over day 0). The percentage of (C) A11 (oligomer)-positive islets, (D) thioflavin S (amyloid) positive islets, and (E) amyloid area to total islet area. Data are presented as mean ± SEM of four independent studies (4 human islet preparations); n = 15–20 islets per condition in each study. *vs day 0; #vs corresponding non-treated group ( p

    Journal: Molecular Metabolism

    Article Title: Amyloid formation disrupts the balance between interleukin-1β and interleukin-1 receptor antagonist in human islets

    doi: 10.1016/j.molmet.2017.05.016

    Figure Lengend Snippet: Treatment with neutralizing IL-1β antibody markedly reduces islet IL-1β immunoreactivity and prevents β-cell Fas upregulation induced by amyloid formation in cultured human islets. Human islets non-transduced or transduced with Ad-prohIAPP-siRNA or Ad-control-siRNA (MOI: 20) were cultured with (+nIL1β) or without (-nIL1β) a neutralizing monoclonal human IL-1β antibody (1 μg/ml) in CMRL (11.1 mmol/L glucose) for 7 days. (A) Paraffin-embedded islet sections were immunolabeled for insulin (red), IL-1β or Fas (green) and thioflavin S (Thio S; blue) as indicated. The squares (dashed white lines) denote regions enlarged and depicted as inserts at the bottom right of each image. The top right inserts in each micrograph show immunolabeling for insulin (green) and A11 (red). Scale bar = 50 μm; inserts: ×3 (A11: ×4). (B) The proportion of Fas positive islet β-cells (fold over day 0). The percentage of (C) A11 (oligomer)-positive islets, (D) thioflavin S (amyloid) positive islets, and (E) amyloid area to total islet area. Data are presented as mean ± SEM of four independent studies (4 human islet preparations); n = 15–20 islets per condition in each study. *vs day 0; #vs corresponding non-treated group ( p

    Article Snippet: For thioflavin S staining, sections were incubated with 0.5% (wt/vol.) thioflavin S solution (Sigma–Aldrich, Oakville, ON, Canada) for 5 min at room temperature.

    Techniques: Cell Culture, Transduction, Immunolabeling