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    • ru486  (MedChemExpress)
    94
    MedChemExpress ru486
    ( A ) Calcium overload level was measured by Fluo4-AM (5 μM, green) calcium indicator of the brain and the muscle in 30 days-old, the relative intensity was analyzed by F/F0, F represented the fluorescence intensity. Brain Scale bar, 2.5 μm, Muscle Scale bar, 5 μm. N=20. ( B ) Western blot of the tissue extracts from the whole body in 30 days-old. Quantification of TH and α-synuclein level. 20 flies were tested for each trial (N=3). ( C ) Immunostaining images of the TH neurons (green) in DaGS>SNCA fly brain PPL1 cluster. Scale bar, 2.5 μm. 5 flies were tested for each trial (N=4). Quantification of the number of the TH positive neurons. ( D ) The lifespan changes of Control (No <t>RU486),</t> DaGS>SNCA/+ , DaGS>TLK/+ and DaGS>SNCA/TLK flies. 80-100 flies were tested for each trial (N=3). (E) TMRM (1 uM, red) staining of the fly brain, the fluorescence intensity was analyzed. Scale bar, 2.5 μm. N=20. ( F ) The same as A with genotype listed on the micrographs. The full gels are shown in the source data.
    Ru486, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ru486/product/MedChemExpress
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ru486 - by Bioz Stars, 2023-02
    94/100 stars
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    male athymic nude mice  (Taconic Biosciences)
    94
    Taconic Biosciences male athymic nude mice
    ( A ) Calcium overload level was measured by Fluo4-AM (5 μM, green) calcium indicator of the brain and the muscle in 30 days-old, the relative intensity was analyzed by F/F0, F represented the fluorescence intensity. Brain Scale bar, 2.5 μm, Muscle Scale bar, 5 μm. N=20. ( B ) Western blot of the tissue extracts from the whole body in 30 days-old. Quantification of TH and α-synuclein level. 20 flies were tested for each trial (N=3). ( C ) Immunostaining images of the TH neurons (green) in DaGS>SNCA fly brain PPL1 cluster. Scale bar, 2.5 μm. 5 flies were tested for each trial (N=4). Quantification of the number of the TH positive neurons. ( D ) The lifespan changes of Control (No <t>RU486),</t> DaGS>SNCA/+ , DaGS>TLK/+ and DaGS>SNCA/TLK flies. 80-100 flies were tested for each trial (N=3). (E) TMRM (1 uM, red) staining of the fly brain, the fluorescence intensity was analyzed. Scale bar, 2.5 μm. N=20. ( F ) The same as A with genotype listed on the micrographs. The full gels are shown in the source data.
    Male Athymic Nude Mice, supplied by Taconic Biosciences, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/male athymic nude mice/product/Taconic Biosciences
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    male athymic nude mice - by Bioz Stars, 2023-02
    94/100 stars
    		  Buy from Supplier
    actinomyces bovis atcc 13683  (ATCC)
    86
    ATCC actinomyces bovis atcc 13683
    ( A ) Calcium overload level was measured by Fluo4-AM (5 μM, green) calcium indicator of the brain and the muscle in 30 days-old, the relative intensity was analyzed by F/F0, F represented the fluorescence intensity. Brain Scale bar, 2.5 μm, Muscle Scale bar, 5 μm. N=20. ( B ) Western blot of the tissue extracts from the whole body in 30 days-old. Quantification of TH and α-synuclein level. 20 flies were tested for each trial (N=3). ( C ) Immunostaining images of the TH neurons (green) in DaGS>SNCA fly brain PPL1 cluster. Scale bar, 2.5 μm. 5 flies were tested for each trial (N=4). Quantification of the number of the TH positive neurons. ( D ) The lifespan changes of Control (No <t>RU486),</t> DaGS>SNCA/+ , DaGS>TLK/+ and DaGS>SNCA/TLK flies. 80-100 flies were tested for each trial (N=3). (E) TMRM (1 uM, red) staining of the fly brain, the fluorescence intensity was analyzed. Scale bar, 2.5 μm. N=20. ( F ) The same as A with genotype listed on the micrographs. The full gels are shown in the source data.
    Actinomyces Bovis Atcc 13683, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/actinomyces bovis atcc 13683/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    actinomyces bovis atcc 13683 - by Bioz Stars, 2023-02
    86/100 stars
    		  Buy from Supplier
    actinomyces bovis atcc 13683 t  (ATCC)
    86
    ATCC actinomyces bovis atcc 13683 t
    ( A ) Calcium overload level was measured by Fluo4-AM (5 μM, green) calcium indicator of the brain and the muscle in 30 days-old, the relative intensity was analyzed by F/F0, F represented the fluorescence intensity. Brain Scale bar, 2.5 μm, Muscle Scale bar, 5 μm. N=20. ( B ) Western blot of the tissue extracts from the whole body in 30 days-old. Quantification of TH and α-synuclein level. 20 flies were tested for each trial (N=3). ( C ) Immunostaining images of the TH neurons (green) in DaGS>SNCA fly brain PPL1 cluster. Scale bar, 2.5 μm. 5 flies were tested for each trial (N=4). Quantification of the number of the TH positive neurons. ( D ) The lifespan changes of Control (No <t>RU486),</t> DaGS>SNCA/+ , DaGS>TLK/+ and DaGS>SNCA/TLK flies. 80-100 flies were tested for each trial (N=3). (E) TMRM (1 uM, red) staining of the fly brain, the fluorescence intensity was analyzed. Scale bar, 2.5 μm. N=20. ( F ) The same as A with genotype listed on the micrographs. The full gels are shown in the source data.
    Actinomyces Bovis Atcc 13683 T, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/actinomyces bovis atcc 13683 t/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    actinomyces bovis atcc 13683 t - by Bioz Stars, 2023-02
    86/100 stars
    		  Buy from Supplier
    anti chmp4b  (Proteintech)
    94
    Proteintech anti chmp4b
    A RAW264.7 macrophages were electroporated with EGFP‐Rab8A and treated with 1 mM LLOMe for 30 min. Intracellular distribution of Galectin‐3, <t>CHMP4B</t> or LC3B was visualised by immunofluorescence. Scale bar = 5 μm. B Macrophages were electroporated with EGFP‐Rab8A and RFP‐Galectin‐3. Cells were then treated with 1 mM of LLOMe and monitored by live cell imaging. Snapshots at the indicated time points after LLOMe addition are shown. Scale bar = 5 μm. C RAW264.7 macrophages were treated with 1 mM of LLOMe, and LRRK2, Rab8A, CHMP4B, Galectin‐3 and LC3B foci over time were analysed by high‐content imaging. Data show the mean ± SEM of five biological replicates. D–H RAW264.7 macrophages were pre‐treated with 10 μM BAPTA‐AM for 1 h, and treated with 1 mM LLOMe for 30 min. (D) CHMP4B recruitment was monitored by immunofluorescence and high‐content imaging. Scale bar = 10 μm. (E) Rab8A pT72 phosphorylation was analysed by Western blot. (F) LRRK2 and (G) Rab8A recruitment was monitored by immunofluorescence and high‐content imaging. Scale bar = 10 μm. (H) Quantification of E and F. (D and H) Data represent the mean ± SEM of two to three independent biological experiments. One‐way ANOVA followed by Sidak's multiple comparisons test. ns = non‐significant; ** P ≤ 0.01, * P ≤ 0.05. Source data are available online for this figure.
    Anti Chmp4b, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti chmp4b/product/Proteintech
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti chmp4b - by Bioz Stars, 2023-02
    94/100 stars
    		  Buy from Supplier

    Image Search Results


    ( A ) Calcium overload level was measured by Fluo4-AM (5 μM, green) calcium indicator of the brain and the muscle in 30 days-old, the relative intensity was analyzed by F/F0, F represented the fluorescence intensity. Brain Scale bar, 2.5 μm, Muscle Scale bar, 5 μm. N=20. ( B ) Western blot of the tissue extracts from the whole body in 30 days-old. Quantification of TH and α-synuclein level. 20 flies were tested for each trial (N=3). ( C ) Immunostaining images of the TH neurons (green) in DaGS>SNCA fly brain PPL1 cluster. Scale bar, 2.5 μm. 5 flies were tested for each trial (N=4). Quantification of the number of the TH positive neurons. ( D ) The lifespan changes of Control (No RU486), DaGS>SNCA/+ , DaGS>TLK/+ and DaGS>SNCA/TLK flies. 80-100 flies were tested for each trial (N=3). (E) TMRM (1 uM, red) staining of the fly brain, the fluorescence intensity was analyzed. Scale bar, 2.5 μm. N=20. ( F ) The same as A with genotype listed on the micrographs. The full gels are shown in the source data.

    Journal: bioRxiv

    Article Title: α-synucleinopathy associated calcium overload and autophagy failure is regulated by gain-of-function of Tousled-like kinase

    doi: 10.1101/2022.07.17.500360

    Figure Lengend Snippet: ( A ) Calcium overload level was measured by Fluo4-AM (5 μM, green) calcium indicator of the brain and the muscle in 30 days-old, the relative intensity was analyzed by F/F0, F represented the fluorescence intensity. Brain Scale bar, 2.5 μm, Muscle Scale bar, 5 μm. N=20. ( B ) Western blot of the tissue extracts from the whole body in 30 days-old. Quantification of TH and α-synuclein level. 20 flies were tested for each trial (N=3). ( C ) Immunostaining images of the TH neurons (green) in DaGS>SNCA fly brain PPL1 cluster. Scale bar, 2.5 μm. 5 flies were tested for each trial (N=4). Quantification of the number of the TH positive neurons. ( D ) The lifespan changes of Control (No RU486), DaGS>SNCA/+ , DaGS>TLK/+ and DaGS>SNCA/TLK flies. 80-100 flies were tested for each trial (N=3). (E) TMRM (1 uM, red) staining of the fly brain, the fluorescence intensity was analyzed. Scale bar, 2.5 μm. N=20. ( F ) The same as A with genotype listed on the micrographs. The full gels are shown in the source data.

    Article Snippet: For DaGS>SNCA flies, 20 flies were collected and placed in one tube at 1-3 days, 500 μM RU486 (Mifepristone; MedChemExpress #HY-13683) was mixed in the standard food, the fly vials were changed to new vials every two days.

    Techniques: Fluorescence, Western Blot, Immunostaining, Staining

    A RAW264.7 macrophages were electroporated with EGFP‐Rab8A and treated with 1 mM LLOMe for 30 min. Intracellular distribution of Galectin‐3, CHMP4B or LC3B was visualised by immunofluorescence. Scale bar = 5 μm. B Macrophages were electroporated with EGFP‐Rab8A and RFP‐Galectin‐3. Cells were then treated with 1 mM of LLOMe and monitored by live cell imaging. Snapshots at the indicated time points after LLOMe addition are shown. Scale bar = 5 μm. C RAW264.7 macrophages were treated with 1 mM of LLOMe, and LRRK2, Rab8A, CHMP4B, Galectin‐3 and LC3B foci over time were analysed by high‐content imaging. Data show the mean ± SEM of five biological replicates. D–H RAW264.7 macrophages were pre‐treated with 10 μM BAPTA‐AM for 1 h, and treated with 1 mM LLOMe for 30 min. (D) CHMP4B recruitment was monitored by immunofluorescence and high‐content imaging. Scale bar = 10 μm. (E) Rab8A pT72 phosphorylation was analysed by Western blot. (F) LRRK2 and (G) Rab8A recruitment was monitored by immunofluorescence and high‐content imaging. Scale bar = 10 μm. (H) Quantification of E and F. (D and H) Data represent the mean ± SEM of two to three independent biological experiments. One‐way ANOVA followed by Sidak's multiple comparisons test. ns = non‐significant; ** P ≤ 0.01, * P ≤ 0.05. Source data are available online for this figure.

    Journal: The EMBO Journal

    Article Title: LRRK 2 activation controls the repair of damaged endomembranes in macrophages

    doi: 10.15252/embj.2020104494

    Figure Lengend Snippet: A RAW264.7 macrophages were electroporated with EGFP‐Rab8A and treated with 1 mM LLOMe for 30 min. Intracellular distribution of Galectin‐3, CHMP4B or LC3B was visualised by immunofluorescence. Scale bar = 5 μm. B Macrophages were electroporated with EGFP‐Rab8A and RFP‐Galectin‐3. Cells were then treated with 1 mM of LLOMe and monitored by live cell imaging. Snapshots at the indicated time points after LLOMe addition are shown. Scale bar = 5 μm. C RAW264.7 macrophages were treated with 1 mM of LLOMe, and LRRK2, Rab8A, CHMP4B, Galectin‐3 and LC3B foci over time were analysed by high‐content imaging. Data show the mean ± SEM of five biological replicates. D–H RAW264.7 macrophages were pre‐treated with 10 μM BAPTA‐AM for 1 h, and treated with 1 mM LLOMe for 30 min. (D) CHMP4B recruitment was monitored by immunofluorescence and high‐content imaging. Scale bar = 10 μm. (E) Rab8A pT72 phosphorylation was analysed by Western blot. (F) LRRK2 and (G) Rab8A recruitment was monitored by immunofluorescence and high‐content imaging. Scale bar = 10 μm. (H) Quantification of E and F. (D and H) Data represent the mean ± SEM of two to three independent biological experiments. One‐way ANOVA followed by Sidak's multiple comparisons test. ns = non‐significant; ** P ≤ 0.01, * P ≤ 0.05. Source data are available online for this figure.

    Article Snippet: Antibodies used in this study were anti‐Rab8A (6975), anti‐Rab10 (8127) and anti‐β‐actin‐HRP (12262) from Cell Signaling; anti‐Rab8A pT72 (ab230260), anti‐Rab10 pT73 (ab230261), anti‐LRRK2 for immunofluorescence (ab133474), anti‐Listeria‐FITC (ab68592), anti‐Candida‐FITC (ab21164) and anti‐LAMP1 for Western Blot (ab24170) from Abcam; anti‐LRRK2 for Western Blot (N241A/34) from NeuroMab; anti‐Galectin‐3‐AF647 (125408) from BioLegend; anti‐CHMP4B (13683‐1AP) from ProteinTech; anti‐LC3B (PM036) from MBL; anti‐Galectin‐8 (AF1305) from R&D Systems; and anti‐LAMP‐1 for immunofluorescence (1D4B) from DSHB and anti‐K63 Ubiquitin (05‐1308) from Millipore.

    Techniques: Immunofluorescence, Live Cell Imaging, Imaging, Western Blot

    RAW264.7 macrophages were electroporated with EGFP‐Rab3A, EGFP‐Rab8A, EGFP‐Rab10 and EGFP‐Rab35. Cells were treated with 1 mM of LLOMe for 30 min, and Rab recruitment to LAMP1 + compartments was monitored by high‐content immunofluorescence imaging. Scale bar = 10 μm. WT and LRRK2 KO RAW264.7 macrophages were treated with 1 mM LLOMe for 30 min. Cells were separated into cytosolic (C) and membrane (M) fractions and analysed for Rab8A pT72, Rab8A and Rab10 by Western blot. WT and Rab8A KO RAW264.7 macrophages were treated with 1 mM LLOMe for 30 min, and Rab8A and Rab8A pT72 levels were analysed by Western blot. RAW264.7 macrophages were electroporated with EGFP‐Rab8A‐WT, EGFP‐Rab8A‐Q67L, EGFP‐Rab8A‐T22N and EGFP‐Rab8A‐T72A. Cells were treated with 1 mM of LLOMe for 30 min, and CHMP4B recruitment was assessed by confocal microscopy. Scale bar = 5 μm. CHMP4B integrated fluorescence density was analysed per cell. Data show values from single cells and mean. Source data are available online for this figure.

    Journal: The EMBO Journal

    Article Title: LRRK 2 activation controls the repair of damaged endomembranes in macrophages

    doi: 10.15252/embj.2020104494

    Figure Lengend Snippet: RAW264.7 macrophages were electroporated with EGFP‐Rab3A, EGFP‐Rab8A, EGFP‐Rab10 and EGFP‐Rab35. Cells were treated with 1 mM of LLOMe for 30 min, and Rab recruitment to LAMP1 + compartments was monitored by high‐content immunofluorescence imaging. Scale bar = 10 μm. WT and LRRK2 KO RAW264.7 macrophages were treated with 1 mM LLOMe for 30 min. Cells were separated into cytosolic (C) and membrane (M) fractions and analysed for Rab8A pT72, Rab8A and Rab10 by Western blot. WT and Rab8A KO RAW264.7 macrophages were treated with 1 mM LLOMe for 30 min, and Rab8A and Rab8A pT72 levels were analysed by Western blot. RAW264.7 macrophages were electroporated with EGFP‐Rab8A‐WT, EGFP‐Rab8A‐Q67L, EGFP‐Rab8A‐T22N and EGFP‐Rab8A‐T72A. Cells were treated with 1 mM of LLOMe for 30 min, and CHMP4B recruitment was assessed by confocal microscopy. Scale bar = 5 μm. CHMP4B integrated fluorescence density was analysed per cell. Data show values from single cells and mean. Source data are available online for this figure.

    Article Snippet: Antibodies used in this study were anti‐Rab8A (6975), anti‐Rab10 (8127) and anti‐β‐actin‐HRP (12262) from Cell Signaling; anti‐Rab8A pT72 (ab230260), anti‐Rab10 pT73 (ab230261), anti‐LRRK2 for immunofluorescence (ab133474), anti‐Listeria‐FITC (ab68592), anti‐Candida‐FITC (ab21164) and anti‐LAMP1 for Western Blot (ab24170) from Abcam; anti‐LRRK2 for Western Blot (N241A/34) from NeuroMab; anti‐Galectin‐3‐AF647 (125408) from BioLegend; anti‐CHMP4B (13683‐1AP) from ProteinTech; anti‐LC3B (PM036) from MBL; anti‐Galectin‐8 (AF1305) from R&D Systems; and anti‐LAMP‐1 for immunofluorescence (1D4B) from DSHB and anti‐K63 Ubiquitin (05‐1308) from Millipore.

    Techniques: Immunofluorescence, Imaging, Western Blot, Confocal Microscopy, Fluorescence

    A, B RAW264.7 WT, LRRK2 KO or Rab8A macrophages pre‐treated with 1 μM GSK2578215A (GSK inh) were treated with 1 mM LLOMe for 30 min. (A) CHMP4B and (B) Galectin‐3 vesicle numbers were analysed by immunofluorescence and high‐content imaging. Scale bar = 20 μm. Right panels show the quantification of number of CHMP4B‐ or Galectin‐3‐positive vesicles per cell. Mean ± SEM of three independent biological experiments. ns = non‐significant, * P ≤ 0.05, ** P ≤ 0.01 by one‐way ANOVA followed by Sidak's multiple comparisons test. C, D Live cell imaging of LysoTracker‐positive spots in WT, LRRK2 KO or Rab8A KO macrophages treated with 1 mM LLOMe, followed by lysosomal recovery after LLOMe wash‐out. One representative experiment out of three shown. Differences between slopes in the LLOMe treatment window were estimated using linear regression.

    Journal: The EMBO Journal

    Article Title: LRRK 2 activation controls the repair of damaged endomembranes in macrophages

    doi: 10.15252/embj.2020104494

    Figure Lengend Snippet: A, B RAW264.7 WT, LRRK2 KO or Rab8A macrophages pre‐treated with 1 μM GSK2578215A (GSK inh) were treated with 1 mM LLOMe for 30 min. (A) CHMP4B and (B) Galectin‐3 vesicle numbers were analysed by immunofluorescence and high‐content imaging. Scale bar = 20 μm. Right panels show the quantification of number of CHMP4B‐ or Galectin‐3‐positive vesicles per cell. Mean ± SEM of three independent biological experiments. ns = non‐significant, * P ≤ 0.05, ** P ≤ 0.01 by one‐way ANOVA followed by Sidak's multiple comparisons test. C, D Live cell imaging of LysoTracker‐positive spots in WT, LRRK2 KO or Rab8A KO macrophages treated with 1 mM LLOMe, followed by lysosomal recovery after LLOMe wash‐out. One representative experiment out of three shown. Differences between slopes in the LLOMe treatment window were estimated using linear regression.

    Article Snippet: Antibodies used in this study were anti‐Rab8A (6975), anti‐Rab10 (8127) and anti‐β‐actin‐HRP (12262) from Cell Signaling; anti‐Rab8A pT72 (ab230260), anti‐Rab10 pT73 (ab230261), anti‐LRRK2 for immunofluorescence (ab133474), anti‐Listeria‐FITC (ab68592), anti‐Candida‐FITC (ab21164) and anti‐LAMP1 for Western Blot (ab24170) from Abcam; anti‐LRRK2 for Western Blot (N241A/34) from NeuroMab; anti‐Galectin‐3‐AF647 (125408) from BioLegend; anti‐CHMP4B (13683‐1AP) from ProteinTech; anti‐LC3B (PM036) from MBL; anti‐Galectin‐8 (AF1305) from R&D Systems; and anti‐LAMP‐1 for immunofluorescence (1D4B) from DSHB and anti‐K63 Ubiquitin (05‐1308) from Millipore.

    Techniques: Immunofluorescence, Imaging, Live Cell Imaging