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Image Search Results
Journal: Oncotarget
Article Title: Extracellular truncated tau causes early presynaptic dysfunction associated with Alzheimer’s disease and other tauopathies
doi: 10.18632/oncotarget.17371
Figure Lengend Snippet: a. - b. Western blotting analysis (n=12) was carried out on equal amounts of total protein extract (40µg) from mature hippocampal primary neurons (DIV15) exposed for 48h to increasing subtoxic concentration (1-2µM) of NH 2 htau and its reverse control sequence. Immunoblots ( a ) were probed with antibodies against several presynaptic- (α-synuclein, synapsin I, synaptosomal-associated protein 25 SNAP-25, synaptophysin, vesicular glutamate transporter 1 vGLUT1 , synaptic vesicle protein 2 SV2, dynamin, synaptotagmin) and post-synaptic markers (N-Methyl-D-aspartate NMDA Receptor Subunit NR1, postsynaptic density protein 95 PSD95) and against not-synaptic proteins located in trans-Golgi network and endoplasmic reticulum (golgin-97 and calnexin). Cropped representative WB are shown. Densitometric quantification of immunoreactivity levels ( b ) was calculated by normalizing the values on the β-actin intensity and expressed as ratio respect to corresponding ctrl values.Values are means of at least nine independent experiments and statistically significant differences were calculated by one-way ANOVA followed by Bonferroni post-hoc test (*p < 0,05; **p<0,01; ***p<0,0001 versus untreated ctrl).
Article Snippet: The following antibodies were used: β-III tubulin antibody rabbit ab18207 Abcam; β-III tubulin antibody mouse (clone 2G10) T8578 Sigma-Aldrich; β-actin mouse S3062 Sigma-Aldrich; Synapsin I antibody rabbit AB1543P Millipore Corporation; PSD95 antibody (clone 7E3-1B8) mouse MAB1598 Millipore; PSD95antibody (clone: 6G6-1C9) mouse ADI-VAM-PS002 EnzoLife Science; synaptophysin antibody mouse sc-17750 Santa Cruz; Calnexin antibody ( clone C-20) goat sc-6465 Santa Cruz ; MAP-2 antibody mouse MAB3418 Millipore; dynamin antibody mouse BD Transduction Laboraories 610246; GAPDH antibody (clone GAPDH-71.1) G8795Sigma-Aldrich ; SNAP25 antibody mouse (clone SMI81) Biolegend 836301; SV2A antibody (clone E-8) mouse sc-376234 Santa Cruz;
Techniques: Western Blot, Concentration Assay, Sequencing
Journal: Oncotarget
Article Title: Extracellular truncated tau causes early presynaptic dysfunction associated with Alzheimer’s disease and other tauopathies
doi: 10.18632/oncotarget.17371
Figure Lengend Snippet: a. - b. - c. Confocal microscopy analysis of double immunofluorescence carried out on mature hippocampal primary neurons (DIV15) exposed for 48h to NH 2 htau and its reverse control sequence (1µM). Merge images show the overlay of the three fluorescence channels, Differential Interference Contrast (DIC; gray channel) enables the visualization of the neuritic network, DIC Merge is the composition of the three fluorescence and of the DIC channels. (a): presynaptic synaptophysin (green channel) and dendritic MAP-2 (red channel). Nuclei (blue) were stained with Hoechst 33258 (0.5 mg/ml). Arrowheads and arrows point to MAP2- positive neurites of larger and smaller caliber, respectively. (b): presynaptic α-synuclein (green channel) and neuron-specific cytoskeletal beta III tubulin (red channel). Arrowheads and arrows point to beta III tubulin-positive neurites of larger and smaller caliber, respectively. (c): presynaptic synapsin I (green channel) and mitochondrial marker COX I (red channel). Arrowheads point to synapsin I-labeled presynaptic spots and arrows point to COX I -positive mitochondrial structures. In the merge, DIC and DIC-Merge channels, arrowheads and arrows appear in opposition to give evidence to mitochondria resident at juxstaposed presynaptic sites. Asterisks mark typical punctuate structures immunoreactive for both synapsin I and COX I (yellow dots) representing mitochondria which are localized to presynaptic sites (i.e.synaptic mitochondria). Note the loss of double-stained synapsin I/COX I puncta and the decrease of juxstaposed presynaptic sites/mitochondria in the NH 2 htau-treated cultures. Images are representative of at least three independent experiments. Scale bar: A=20 µm ;B-C=10 µm.
Article Snippet: The following antibodies were used: β-III tubulin antibody rabbit ab18207 Abcam; β-III tubulin antibody mouse (clone 2G10) T8578 Sigma-Aldrich; β-actin mouse S3062 Sigma-Aldrich; Synapsin I antibody rabbit AB1543P Millipore Corporation; PSD95 antibody (clone 7E3-1B8) mouse MAB1598 Millipore; PSD95antibody (clone: 6G6-1C9) mouse ADI-VAM-PS002 EnzoLife Science; synaptophysin antibody mouse sc-17750 Santa Cruz; Calnexin antibody ( clone C-20) goat sc-6465 Santa Cruz ; MAP-2 antibody mouse MAB3418 Millipore; dynamin antibody mouse BD Transduction Laboraories 610246; GAPDH antibody (clone GAPDH-71.1) G8795Sigma-Aldrich ; SNAP25 antibody mouse (clone SMI81) Biolegend 836301; SV2A antibody (clone E-8) mouse sc-376234 Santa Cruz;
Techniques: Confocal Microscopy, Immunofluorescence, Sequencing, Fluorescence, Staining, Marker, Labeling
Journal: Oncotarget
Article Title: Extracellular truncated tau causes early presynaptic dysfunction associated with Alzheimer's disease and other tauopathies.
doi: 10.18632/oncotarget.17371
Figure Lengend Snippet: Figure 5: Long-term application of NH2htau induces a marked and selective loss of exocytotic presynaptic vesicles proteins in cultured hippocampal neurons. a.-b. Western blotting analysis (n=12) was carried out on equal amounts of total protein extract (40µg) from mature hippocampal primary neurons (DIV15) exposed for 48h to increasing subtoxic concentration (1-2µM) of NH2htau and its reverse control sequence. Immunoblots (a) were probed with antibodies against several presynaptic- (α-synuclein, synapsin I, synaptosomal-associated protein 25 SNAP-25, synaptophysin, vesicular glutamate transporter 1 vGLUT1 , synaptic vesicle protein 2 SV2, dynamin, synaptotagmin) and post-synaptic markers (N-Methyl-D-aspartate NMDA Receptor Subunit NR1, postsynaptic density protein 95 PSD95) and against not-synaptic proteins located in trans-Golgi network and endoplasmic reticulum (golgin-97 and calnexin). Cropped representative WB are shown. Densitometric quantification of immunoreactivity levels (b) was calculated by normalizing the values on the β-actin intensity and expressed as ratio respect to corresponding ctrl values.Values are means of at least nine independent experiments and statistically significant differences were calculated by one-way ANOVA followed by Bonferroni post-hoc test (*p < 0,05; **p<0,01; ***p<0,0001 versus untreated ctrl).
Article Snippet: The following antibodies were used: β-III tubulin antibody rabbit ab18207 Abcam; β-III tubulin antibody mouse (clone 2G10) T8578 Sigma-Aldrich; β-actin mouse S3062 Sigma-Aldrich; Synapsin I antibody rabbit AB1543P Millipore Corporation; PSD95 antibody (clone 7E3-1B8) mouse MAB1598 Millipore; PSD95antibody (clone: 6G6-1C9) mouse ADI-VAM-PS002 EnzoLife Science; synaptophysin antibody mouse sc-17750 Santa Cruz;
Techniques: Cell Culture, Western Blot, Concentration Assay, Control, Sequencing