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stx7 antibody  (Bethyl)


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    Structured Review

    Bethyl stx7 antibody
    Figure 4. DEP impair autophagic flux in the degradation stage. (a) HUVECs infected with RFP-LC3B-GFP- expressing adenovirus were exposed to DEP (70 µg/ml) for 18 h and autolysosome formation was examined under fluorescence microscopy. (b) The number of yellow signals per cell was counted from 10 randomly selected cells. Statistical analysis was performed using two-tailed Student’s t test. **P < 0.02 versus control. (c) HUVECs were pre-incubated with Bafilomysin A1 (Baf A1, 100 nM) for 3 h and exposed to DEP (70 μg/ ml) for an additional 12 h. LC3B level was measured by immunoblotting. (d) Quantification of LC3BII levels normalized to β-actin. Results are presented as means ± SD (n = 3). Statistical analysis was performed using one-way ANOVA. **P < 0.02 versus no treatment. (e) HUVECs were exposed to DEP (70 μg/ml) for 24 h and protein levels of STX17, VAMP8, SNAP29, YKT6, <t>STX7,</t> and β-actin were measured by immunoblotting. (f) Quantification of STX17, VAMP8, SNAP29, YKT6, and STX7 levels normalized to β-actin. Results are presented as means ± SD (n = 3). Statistical analysis was performed using two-tailed Student’s t test. **P < 0.02 versus control.
    Stx7 Antibody, supplied by Bethyl, used in various techniques. Bioz Stars score: 93/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/a304+512a/Syntaxin+7+Antibody/pm36192481-233-1-6
    Average 93 stars, based on 14 article reviews
    stx7 antibody - by Bioz Stars, 2026-10
    93/100 stars

    Images

    1) Product Images from "Diesel exhaust particles induce human umbilical vein endothelial cells apoptosis by accumulation of autophagosomes and caspase-8 activation."

    Article Title: Diesel exhaust particles induce human umbilical vein endothelial cells apoptosis by accumulation of autophagosomes and caspase-8 activation.

    Journal: Scientific reports

    doi: 10.1038/s41598-022-21044-3

    Figure 4. DEP impair autophagic flux in the degradation stage. (a) HUVECs infected with RFP-LC3B-GFP- expressing adenovirus were exposed to DEP (70 µg/ml) for 18 h and autolysosome formation was examined under fluorescence microscopy. (b) The number of yellow signals per cell was counted from 10 randomly selected cells. Statistical analysis was performed using two-tailed Student’s t test. **P < 0.02 versus control. (c) HUVECs were pre-incubated with Bafilomysin A1 (Baf A1, 100 nM) for 3 h and exposed to DEP (70 μg/ ml) for an additional 12 h. LC3B level was measured by immunoblotting. (d) Quantification of LC3BII levels normalized to β-actin. Results are presented as means ± SD (n = 3). Statistical analysis was performed using one-way ANOVA. **P < 0.02 versus no treatment. (e) HUVECs were exposed to DEP (70 μg/ml) for 24 h and protein levels of STX17, VAMP8, SNAP29, YKT6, STX7, and β-actin were measured by immunoblotting. (f) Quantification of STX17, VAMP8, SNAP29, YKT6, and STX7 levels normalized to β-actin. Results are presented as means ± SD (n = 3). Statistical analysis was performed using two-tailed Student’s t test. **P < 0.02 versus control.
    Figure Legend Snippet: Figure 4. DEP impair autophagic flux in the degradation stage. (a) HUVECs infected with RFP-LC3B-GFP- expressing adenovirus were exposed to DEP (70 µg/ml) for 18 h and autolysosome formation was examined under fluorescence microscopy. (b) The number of yellow signals per cell was counted from 10 randomly selected cells. Statistical analysis was performed using two-tailed Student’s t test. **P < 0.02 versus control. (c) HUVECs were pre-incubated with Bafilomysin A1 (Baf A1, 100 nM) for 3 h and exposed to DEP (70 μg/ ml) for an additional 12 h. LC3B level was measured by immunoblotting. (d) Quantification of LC3BII levels normalized to β-actin. Results are presented as means ± SD (n = 3). Statistical analysis was performed using one-way ANOVA. **P < 0.02 versus no treatment. (e) HUVECs were exposed to DEP (70 μg/ml) for 24 h and protein levels of STX17, VAMP8, SNAP29, YKT6, STX7, and β-actin were measured by immunoblotting. (f) Quantification of STX17, VAMP8, SNAP29, YKT6, and STX7 levels normalized to β-actin. Results are presented as means ± SD (n = 3). Statistical analysis was performed using two-tailed Student’s t test. **P < 0.02 versus control.

    Techniques Used: Infection, Expressing, Fluorescence, Microscopy, Two Tailed Test, Control, Incubation, Western Blot

    Related Articles

    Western Blot:

    Article Title: TNF-induced necroptosis initiates early autophagy events via RIPK3-dependent AMPK activation, but inhibits late autophagy
    Article Snippet: .. Anti-mouse RIPK3 (ProSci, 2283), anti-MLKL phospho-Ser345 [EPR9515(2)] (Abcam, ab196436), anti-MLKL (Sigma-Aldrich, SAB1302339), anti-FLAG (Sigma-Aldrich, F3165), anti-ACTB/β-actin (Sigma-Aldrich, A5316), anti-PRKAA1/2 (Cell Signaling Technology, 2793 (clone F6) and 2603 (clone 23A3) for immunoblot, and Santa Cruz Biotechnology, sc-74,461 (clone D-6) for immunopurification), anti-PRKAA1/2 phospho-T183/T172 (40H9) (Cell Signaling Technology, 2535), anti-GST (Sigma-Aldrich, G7781), anti-LC3B (Cell Signaling Technology, 2775 for immunoblot [detects endogenous levels of total LC3B protein; cross-reactivity may exist with other LC3 isoforms according to manufacturer specification], and MBL International; PM036 for immunofluorescence [reacts with LC3A/LC3B/LC3C according to manufacturer specification]), anti-BECN1 (H-300) (Santa Cruz Biotechnology, sc-11,427), anti-BECN1 phospho-S93 (D9A5G) (Cell Signaling Technology, 14717S), anti-ATG14 (Santa Cruz Biotechnology, sc-164767), anti-ACACA phospho-S79 (Cell Signaling Technology, 3661), anti-ACACA (Cell Signaling Technology, 3662), anti-ULK1 phospho-S555 (Cell Signaling Technology, 5869), anti-ULK1 (Cell Signaling Technology, 8054), anti-ATG16L1 (MBL International, PM040), anti-STX17 (Sigma-Aldrich, HPA001204), anti-TAX1BP1 (Proteintech, 14424-1-AP), anti-anti-SQSTM1/p62 (C terminus) (PROGEN, GP62-C), anti-TUBA1A/tubulin (Sigma-Aldrich, T5168), anti-GAPDH (Abcam, ab8245), anti-CHMP4B (Cell Signaling Technology, 42466), anti-SNAP29 [EPR9198(2)] (Abcam, ab181151), anti-VAMP8 (Cell Signaling Technology, 13060), anti-STX7 (Bethyl Laboratories, A304-512A-M), anti-YKT6 (Santa Cruz Biotechnology, sc-365732), anti-GFP (Nacalai USA, 04404–84), anti-ATG13 (MBL International, M183-3). .. IRDye 800- or IRDye 680-conjugated secondary antibodies were purchased from LI-COR Biosciences (925–32212/13/14, 925–68072/73/74).

    Article Title: Kansl1 haploinsufficiency impairs autophagosome-lysosome fusion and links autophagic dysfunction with Koolen-de Vries syndrome in mice.
    Article Snippet: Cells were collected and resuspended in RIPA lysis buffer (50 mM Tris, pH 7.4, 150 mM NaCl, 1% NP-40 [Santa Cruz Biotechnology, CAS 9016–45-9], 0.5% Na-deoxycholate [Sigma-Aldrich, D6750]) containing certain protease inhibitors (0.1 mM phenylmethanesulfonylfluoride [Target Molecule Corporation, T0789], 1× EDTA-free Protease Inhibitor Cocktail [ROCHE, 11873580001]) on ice. .. Antibodies were used at the following concentrations: rabbit anti-KANSL1 (Abnova, PAB20355; 1:1000) for KANSL1 (Homo sapiens) (Note: this antibody has been purified with the PVDF membrane blotted with Kansl1 knockout MEFs samples before use.), rabbit anti-Kansl1 (AtaGenix, fragments of Kansl1 814-1036 aa were used to immunize rats) for Kansl1 (Mus musculus), rabbit anti-STX17 (Sigma, HPA001204; 1:500), rabbit anti-ATG14 (CST, 5504S, 1:500), rabbit anti-PIK3R4 (CST, 14580S, 1:1000), rabbit anti-VAMP8 (CST, 13060S, 1:500), mouse anti-YKT6 (Santa Cruz Biotechnology, sc-365732, 1:1000), rabbit anti-STX7 (Bethyl, A304-512A, 1:2000), rabbit anti-SNAP29 (Proteintech, 12704-1-AP, 1:1000), rabbit anti-MOF (Bethyl, A300-994A; 1:500), rabbit antiLC3B (Sigma, l7543; 1:5000), rabbit-p62 (MBL, PM045; 1:5000), rabbit antiGAPDH (1:5000) prepared in our laboratory and generated by immunizing rabbits with human GAPDH protein, mouse anti-TUBA4A/tubulin (Sigma-Aldrich, T5168; 1:5000), and total OXPHOS Rodent WB Antibody Cocktail (Abcam, ab110413, 1:1000) incubated for 1 h at room temperature or overnight at 4 °C. .. Membranes were incubated with HRP-conjugated secondary antibodies (Dako Cytomation, P0447 and P0448, 1:5000) for 1 h at room temperature.

    Article Title: A conformational switch driven by phosphorylation regulates the activity of the evolutionarily conserved SNARE Ykt6
    Article Snippet: Secondary antibodies used are the following: Alexa 488 (Invitrogen, a21202 and a21206), Alexa 594 (Invitrogen, a21442) and Alexa 647 (Invitrogen, a281883). .. For Western Blot the following primary antibodies were used: GFP (Santa Cruz, sc-9996), actin (Abcam, ab6276), Ykt6 (SantaCruz, sc-365732), LC3B (Cell Signaling, #2775), p62 (Sigma, p0067), STX17 (Sigma, hpa001204), SNAP29 (Abcam, ab138500) Na+/K+ ATPase (Sigma, 06-172-I), alpha/beta-Tubulin (Cell Signaling, 2148S), and STX7 (Bethyl, A304-512A). ..

    Article Title: PACSIN1 is indispensable for amphisome-lysosome fusion during basal autophagy and subsets of selective autophagy.
    Article Snippet: The following reagents were purchased from the indicated companies; Baf.A1 (wako, 029– 11643), E-64-d (Peptide Institute, 4321-v), and pepstatin A (Peptide Institute, 4397-v), MG132 (Miilipore, 474790), puromycin (Invitrogen, ant-pr-1). .. For immunoblotting, the following antibodies were used at the indicated dilutions: antiLC3 (rabbit, 1/1500; MBL, pM036), anti-p62 (rabbit, 1/5000; MBL, pM045), anti–α-tubulin (mouse, 1/50000; Sigma-Aldrich, T5168), anti-ATG13 (rabbit, 1/1000; Sigma-Aldrich, SAB4200100), anti-SNAP29 (rabbit, 1/1000; Abcam, 138500 (EPR9199)), anti-VAMP8 (rabbit, 1/10000; Abcam, 76021), anti-STX7 (rabbit, 1/1000; Bethyl, A304-512A-T), anti-GFP (rabbit, 1/1000; MBL, 598), anti-UQCRC1 (Complex III core subunit 1) (mouse, 1/2000; Invitrogen, 459140), anti-COXIV (rabbit, 1/1000; CST, 4850 (3E11)), anti-GFP (mouse, 1/2000; Santa Cruz Biotechnology, 9996 (B-2)), anti-PACSINs (rabbit, 1/1000; [23]), anti-PACSIN3 (mouse, 1/100; Santa Cruz Biotechnology, 373952 (F-8)), anti-Rab7 (mouse, 1/1000; CST, 95746s (E907E)), anti-GABARAP (rabbit, 1/1000; MBL, pM037), anti-Vps41 (mouse, 1/100; Santa Cruz Biotechnology, 377118 (D-12)), anti-Vps39 (mouse, 1/100; Santa Cruz Biotechnology, 514762 (C-5)), anti-STX17 (rabbit, 1/1000; Sigma-Aldrich, HPA001204), anti-YKT6p (mouse, 1/100; Santa Cruz Biotechnology, 365732 (E-2)), anti-FK2 (mouse, 1/1000; Nippon Bio-test Laboratories, 0918–2), anti-Lamin B (goat, 1/1000; Santa Cruz Biotechnology, 6217(M-20)), and anti-FLAG (rabbit, 1/2000; Sigma-Aldrich, F7425). .. The secondary antibodies used for immunoblotting were as follows: HRP-conjugated goat anti–rabbit IgG (1/5000; Jackson ImmunoResearch, 111-035-003) and HRP-conjugated goat anti–mouse IgG (1/5000; Jackson ImmunoResearch, 115-035-003).

    Immu-Puri:

    Article Title: TNF-induced necroptosis initiates early autophagy events via RIPK3-dependent AMPK activation, but inhibits late autophagy
    Article Snippet: .. Anti-mouse RIPK3 (ProSci, 2283), anti-MLKL phospho-Ser345 [EPR9515(2)] (Abcam, ab196436), anti-MLKL (Sigma-Aldrich, SAB1302339), anti-FLAG (Sigma-Aldrich, F3165), anti-ACTB/β-actin (Sigma-Aldrich, A5316), anti-PRKAA1/2 (Cell Signaling Technology, 2793 (clone F6) and 2603 (clone 23A3) for immunoblot, and Santa Cruz Biotechnology, sc-74,461 (clone D-6) for immunopurification), anti-PRKAA1/2 phospho-T183/T172 (40H9) (Cell Signaling Technology, 2535), anti-GST (Sigma-Aldrich, G7781), anti-LC3B (Cell Signaling Technology, 2775 for immunoblot [detects endogenous levels of total LC3B protein; cross-reactivity may exist with other LC3 isoforms according to manufacturer specification], and MBL International; PM036 for immunofluorescence [reacts with LC3A/LC3B/LC3C according to manufacturer specification]), anti-BECN1 (H-300) (Santa Cruz Biotechnology, sc-11,427), anti-BECN1 phospho-S93 (D9A5G) (Cell Signaling Technology, 14717S), anti-ATG14 (Santa Cruz Biotechnology, sc-164767), anti-ACACA phospho-S79 (Cell Signaling Technology, 3661), anti-ACACA (Cell Signaling Technology, 3662), anti-ULK1 phospho-S555 (Cell Signaling Technology, 5869), anti-ULK1 (Cell Signaling Technology, 8054), anti-ATG16L1 (MBL International, PM040), anti-STX17 (Sigma-Aldrich, HPA001204), anti-TAX1BP1 (Proteintech, 14424-1-AP), anti-anti-SQSTM1/p62 (C terminus) (PROGEN, GP62-C), anti-TUBA1A/tubulin (Sigma-Aldrich, T5168), anti-GAPDH (Abcam, ab8245), anti-CHMP4B (Cell Signaling Technology, 42466), anti-SNAP29 [EPR9198(2)] (Abcam, ab181151), anti-VAMP8 (Cell Signaling Technology, 13060), anti-STX7 (Bethyl Laboratories, A304-512A-M), anti-YKT6 (Santa Cruz Biotechnology, sc-365732), anti-GFP (Nacalai USA, 04404–84), anti-ATG13 (MBL International, M183-3). .. IRDye 800- or IRDye 680-conjugated secondary antibodies were purchased from LI-COR Biosciences (925–32212/13/14, 925–68072/73/74).

    Immunofluorescence:

    Article Title: TNF-induced necroptosis initiates early autophagy events via RIPK3-dependent AMPK activation, but inhibits late autophagy
    Article Snippet: .. Anti-mouse RIPK3 (ProSci, 2283), anti-MLKL phospho-Ser345 [EPR9515(2)] (Abcam, ab196436), anti-MLKL (Sigma-Aldrich, SAB1302339), anti-FLAG (Sigma-Aldrich, F3165), anti-ACTB/β-actin (Sigma-Aldrich, A5316), anti-PRKAA1/2 (Cell Signaling Technology, 2793 (clone F6) and 2603 (clone 23A3) for immunoblot, and Santa Cruz Biotechnology, sc-74,461 (clone D-6) for immunopurification), anti-PRKAA1/2 phospho-T183/T172 (40H9) (Cell Signaling Technology, 2535), anti-GST (Sigma-Aldrich, G7781), anti-LC3B (Cell Signaling Technology, 2775 for immunoblot [detects endogenous levels of total LC3B protein; cross-reactivity may exist with other LC3 isoforms according to manufacturer specification], and MBL International; PM036 for immunofluorescence [reacts with LC3A/LC3B/LC3C according to manufacturer specification]), anti-BECN1 (H-300) (Santa Cruz Biotechnology, sc-11,427), anti-BECN1 phospho-S93 (D9A5G) (Cell Signaling Technology, 14717S), anti-ATG14 (Santa Cruz Biotechnology, sc-164767), anti-ACACA phospho-S79 (Cell Signaling Technology, 3661), anti-ACACA (Cell Signaling Technology, 3662), anti-ULK1 phospho-S555 (Cell Signaling Technology, 5869), anti-ULK1 (Cell Signaling Technology, 8054), anti-ATG16L1 (MBL International, PM040), anti-STX17 (Sigma-Aldrich, HPA001204), anti-TAX1BP1 (Proteintech, 14424-1-AP), anti-anti-SQSTM1/p62 (C terminus) (PROGEN, GP62-C), anti-TUBA1A/tubulin (Sigma-Aldrich, T5168), anti-GAPDH (Abcam, ab8245), anti-CHMP4B (Cell Signaling Technology, 42466), anti-SNAP29 [EPR9198(2)] (Abcam, ab181151), anti-VAMP8 (Cell Signaling Technology, 13060), anti-STX7 (Bethyl Laboratories, A304-512A-M), anti-YKT6 (Santa Cruz Biotechnology, sc-365732), anti-GFP (Nacalai USA, 04404–84), anti-ATG13 (MBL International, M183-3). .. IRDye 800- or IRDye 680-conjugated secondary antibodies were purchased from LI-COR Biosciences (925–32212/13/14, 925–68072/73/74).

    Expressing:

    Article Title: The endosomal Q-SNARE, Syntaxin 7, defines a rapidly replenishing synaptic vesicle recycling pool in hippocampal neurons
    Article Snippet: .. Primary Antibodies Anti-Syntaxin-7 (Bethyl Laboratories, A304-512A, Bethyl Laboratories) Anti-GFP (Rabbit polyclonal, kindly gifted from Reinhard Jahn) Anti-Synaptobrevin 2 (Mouse monoclonal, Cl69.1, kindly gifted from Reinhard Jahn) Anti-Synaptophysin (Mouse monoclonal, Cl7.2, kind gifted from Reinhard Jahn) Anti-Synaptophysin (Guinea pig polyclonal, 101 004, Synaptic Systems) Anti-MAP2 (Chick polyclonal, ab5392, Abcam) Anti-Tubulin beta3 (Mouse monoclonal, clone Tuj1, #MMS-435P, Covance) Anti-Bassoon (Mouse monoclonal, SAP7F407, Novus Biologicals) Secondary antibodies Alexa-488-conjugated anti-rabbit IgG (A27034, Invitrogen) Alexa-555-conjugated anti-mouse IgG (A28180, Invitrogen) Alexa-633-conjugated anti-chick IgG (A11039, Invitrogen) Alexa-633-conjugated anti-guinea pig IgG (A21005, Invitrogen) HRP-linked anti-mouse IgG (kindly gifted from Reinhard Jahn) HRP-linked anti-rabbit IgG (NA934-1ML, GE Healthcare) FluoroNanogold-anti-rabbit Fab'-Alexa Fluo 488 (7204, Nanoprobes) The specificity of the Stx7 antibody was confirmed in independent experiments where Stx7 expression was reduced by specific shRNAs (supplementary figure. ..

    Purification:

    Article Title: Kansl1 haploinsufficiency impairs autophagosome-lysosome fusion and links autophagic dysfunction with Koolen-de Vries syndrome in mice.
    Article Snippet: Cells were collected and resuspended in RIPA lysis buffer (50 mM Tris, pH 7.4, 150 mM NaCl, 1% NP-40 [Santa Cruz Biotechnology, CAS 9016–45-9], 0.5% Na-deoxycholate [Sigma-Aldrich, D6750]) containing certain protease inhibitors (0.1 mM phenylmethanesulfonylfluoride [Target Molecule Corporation, T0789], 1× EDTA-free Protease Inhibitor Cocktail [ROCHE, 11873580001]) on ice. .. Antibodies were used at the following concentrations: rabbit anti-KANSL1 (Abnova, PAB20355; 1:1000) for KANSL1 (Homo sapiens) (Note: this antibody has been purified with the PVDF membrane blotted with Kansl1 knockout MEFs samples before use.), rabbit anti-Kansl1 (AtaGenix, fragments of Kansl1 814-1036 aa were used to immunize rats) for Kansl1 (Mus musculus), rabbit anti-STX17 (Sigma, HPA001204; 1:500), rabbit anti-ATG14 (CST, 5504S, 1:500), rabbit anti-PIK3R4 (CST, 14580S, 1:1000), rabbit anti-VAMP8 (CST, 13060S, 1:500), mouse anti-YKT6 (Santa Cruz Biotechnology, sc-365732, 1:1000), rabbit anti-STX7 (Bethyl, A304-512A, 1:2000), rabbit anti-SNAP29 (Proteintech, 12704-1-AP, 1:1000), rabbit anti-MOF (Bethyl, A300-994A; 1:500), rabbit antiLC3B (Sigma, l7543; 1:5000), rabbit-p62 (MBL, PM045; 1:5000), rabbit antiGAPDH (1:5000) prepared in our laboratory and generated by immunizing rabbits with human GAPDH protein, mouse anti-TUBA4A/tubulin (Sigma-Aldrich, T5168; 1:5000), and total OXPHOS Rodent WB Antibody Cocktail (Abcam, ab110413, 1:1000) incubated for 1 h at room temperature or overnight at 4 °C. .. Membranes were incubated with HRP-conjugated secondary antibodies (Dako Cytomation, P0447 and P0448, 1:5000) for 1 h at room temperature.

    Membrane:

    Article Title: Kansl1 haploinsufficiency impairs autophagosome-lysosome fusion and links autophagic dysfunction with Koolen-de Vries syndrome in mice.
    Article Snippet: Cells were collected and resuspended in RIPA lysis buffer (50 mM Tris, pH 7.4, 150 mM NaCl, 1% NP-40 [Santa Cruz Biotechnology, CAS 9016–45-9], 0.5% Na-deoxycholate [Sigma-Aldrich, D6750]) containing certain protease inhibitors (0.1 mM phenylmethanesulfonylfluoride [Target Molecule Corporation, T0789], 1× EDTA-free Protease Inhibitor Cocktail [ROCHE, 11873580001]) on ice. .. Antibodies were used at the following concentrations: rabbit anti-KANSL1 (Abnova, PAB20355; 1:1000) for KANSL1 (Homo sapiens) (Note: this antibody has been purified with the PVDF membrane blotted with Kansl1 knockout MEFs samples before use.), rabbit anti-Kansl1 (AtaGenix, fragments of Kansl1 814-1036 aa were used to immunize rats) for Kansl1 (Mus musculus), rabbit anti-STX17 (Sigma, HPA001204; 1:500), rabbit anti-ATG14 (CST, 5504S, 1:500), rabbit anti-PIK3R4 (CST, 14580S, 1:1000), rabbit anti-VAMP8 (CST, 13060S, 1:500), mouse anti-YKT6 (Santa Cruz Biotechnology, sc-365732, 1:1000), rabbit anti-STX7 (Bethyl, A304-512A, 1:2000), rabbit anti-SNAP29 (Proteintech, 12704-1-AP, 1:1000), rabbit anti-MOF (Bethyl, A300-994A; 1:500), rabbit antiLC3B (Sigma, l7543; 1:5000), rabbit-p62 (MBL, PM045; 1:5000), rabbit antiGAPDH (1:5000) prepared in our laboratory and generated by immunizing rabbits with human GAPDH protein, mouse anti-TUBA4A/tubulin (Sigma-Aldrich, T5168; 1:5000), and total OXPHOS Rodent WB Antibody Cocktail (Abcam, ab110413, 1:1000) incubated for 1 h at room temperature or overnight at 4 °C. .. Membranes were incubated with HRP-conjugated secondary antibodies (Dako Cytomation, P0447 and P0448, 1:5000) for 1 h at room temperature.

    Knock-Out:

    Article Title: Kansl1 haploinsufficiency impairs autophagosome-lysosome fusion and links autophagic dysfunction with Koolen-de Vries syndrome in mice.
    Article Snippet: Cells were collected and resuspended in RIPA lysis buffer (50 mM Tris, pH 7.4, 150 mM NaCl, 1% NP-40 [Santa Cruz Biotechnology, CAS 9016–45-9], 0.5% Na-deoxycholate [Sigma-Aldrich, D6750]) containing certain protease inhibitors (0.1 mM phenylmethanesulfonylfluoride [Target Molecule Corporation, T0789], 1× EDTA-free Protease Inhibitor Cocktail [ROCHE, 11873580001]) on ice. .. Antibodies were used at the following concentrations: rabbit anti-KANSL1 (Abnova, PAB20355; 1:1000) for KANSL1 (Homo sapiens) (Note: this antibody has been purified with the PVDF membrane blotted with Kansl1 knockout MEFs samples before use.), rabbit anti-Kansl1 (AtaGenix, fragments of Kansl1 814-1036 aa were used to immunize rats) for Kansl1 (Mus musculus), rabbit anti-STX17 (Sigma, HPA001204; 1:500), rabbit anti-ATG14 (CST, 5504S, 1:500), rabbit anti-PIK3R4 (CST, 14580S, 1:1000), rabbit anti-VAMP8 (CST, 13060S, 1:500), mouse anti-YKT6 (Santa Cruz Biotechnology, sc-365732, 1:1000), rabbit anti-STX7 (Bethyl, A304-512A, 1:2000), rabbit anti-SNAP29 (Proteintech, 12704-1-AP, 1:1000), rabbit anti-MOF (Bethyl, A300-994A; 1:500), rabbit antiLC3B (Sigma, l7543; 1:5000), rabbit-p62 (MBL, PM045; 1:5000), rabbit antiGAPDH (1:5000) prepared in our laboratory and generated by immunizing rabbits with human GAPDH protein, mouse anti-TUBA4A/tubulin (Sigma-Aldrich, T5168; 1:5000), and total OXPHOS Rodent WB Antibody Cocktail (Abcam, ab110413, 1:1000) incubated for 1 h at room temperature or overnight at 4 °C. .. Membranes were incubated with HRP-conjugated secondary antibodies (Dako Cytomation, P0447 and P0448, 1:5000) for 1 h at room temperature.

    Generated:

    Article Title: Kansl1 haploinsufficiency impairs autophagosome-lysosome fusion and links autophagic dysfunction with Koolen-de Vries syndrome in mice.
    Article Snippet: Cells were collected and resuspended in RIPA lysis buffer (50 mM Tris, pH 7.4, 150 mM NaCl, 1% NP-40 [Santa Cruz Biotechnology, CAS 9016–45-9], 0.5% Na-deoxycholate [Sigma-Aldrich, D6750]) containing certain protease inhibitors (0.1 mM phenylmethanesulfonylfluoride [Target Molecule Corporation, T0789], 1× EDTA-free Protease Inhibitor Cocktail [ROCHE, 11873580001]) on ice. .. Antibodies were used at the following concentrations: rabbit anti-KANSL1 (Abnova, PAB20355; 1:1000) for KANSL1 (Homo sapiens) (Note: this antibody has been purified with the PVDF membrane blotted with Kansl1 knockout MEFs samples before use.), rabbit anti-Kansl1 (AtaGenix, fragments of Kansl1 814-1036 aa were used to immunize rats) for Kansl1 (Mus musculus), rabbit anti-STX17 (Sigma, HPA001204; 1:500), rabbit anti-ATG14 (CST, 5504S, 1:500), rabbit anti-PIK3R4 (CST, 14580S, 1:1000), rabbit anti-VAMP8 (CST, 13060S, 1:500), mouse anti-YKT6 (Santa Cruz Biotechnology, sc-365732, 1:1000), rabbit anti-STX7 (Bethyl, A304-512A, 1:2000), rabbit anti-SNAP29 (Proteintech, 12704-1-AP, 1:1000), rabbit anti-MOF (Bethyl, A300-994A; 1:500), rabbit antiLC3B (Sigma, l7543; 1:5000), rabbit-p62 (MBL, PM045; 1:5000), rabbit antiGAPDH (1:5000) prepared in our laboratory and generated by immunizing rabbits with human GAPDH protein, mouse anti-TUBA4A/tubulin (Sigma-Aldrich, T5168; 1:5000), and total OXPHOS Rodent WB Antibody Cocktail (Abcam, ab110413, 1:1000) incubated for 1 h at room temperature or overnight at 4 °C. .. Membranes were incubated with HRP-conjugated secondary antibodies (Dako Cytomation, P0447 and P0448, 1:5000) for 1 h at room temperature.

    Incubation:

    Article Title: Kansl1 haploinsufficiency impairs autophagosome-lysosome fusion and links autophagic dysfunction with Koolen-de Vries syndrome in mice.
    Article Snippet: Cells were collected and resuspended in RIPA lysis buffer (50 mM Tris, pH 7.4, 150 mM NaCl, 1% NP-40 [Santa Cruz Biotechnology, CAS 9016–45-9], 0.5% Na-deoxycholate [Sigma-Aldrich, D6750]) containing certain protease inhibitors (0.1 mM phenylmethanesulfonylfluoride [Target Molecule Corporation, T0789], 1× EDTA-free Protease Inhibitor Cocktail [ROCHE, 11873580001]) on ice. .. Antibodies were used at the following concentrations: rabbit anti-KANSL1 (Abnova, PAB20355; 1:1000) for KANSL1 (Homo sapiens) (Note: this antibody has been purified with the PVDF membrane blotted with Kansl1 knockout MEFs samples before use.), rabbit anti-Kansl1 (AtaGenix, fragments of Kansl1 814-1036 aa were used to immunize rats) for Kansl1 (Mus musculus), rabbit anti-STX17 (Sigma, HPA001204; 1:500), rabbit anti-ATG14 (CST, 5504S, 1:500), rabbit anti-PIK3R4 (CST, 14580S, 1:1000), rabbit anti-VAMP8 (CST, 13060S, 1:500), mouse anti-YKT6 (Santa Cruz Biotechnology, sc-365732, 1:1000), rabbit anti-STX7 (Bethyl, A304-512A, 1:2000), rabbit anti-SNAP29 (Proteintech, 12704-1-AP, 1:1000), rabbit anti-MOF (Bethyl, A300-994A; 1:500), rabbit antiLC3B (Sigma, l7543; 1:5000), rabbit-p62 (MBL, PM045; 1:5000), rabbit antiGAPDH (1:5000) prepared in our laboratory and generated by immunizing rabbits with human GAPDH protein, mouse anti-TUBA4A/tubulin (Sigma-Aldrich, T5168; 1:5000), and total OXPHOS Rodent WB Antibody Cocktail (Abcam, ab110413, 1:1000) incubated for 1 h at room temperature or overnight at 4 °C. .. Membranes were incubated with HRP-conjugated secondary antibodies (Dako Cytomation, P0447 and P0448, 1:5000) for 1 h at room temperature.

    other:

    Article Title: Persistent coxsackievirus B1 infection triggers extensive changes in the transcriptome of human pancreatic ductal cells
    Article Snippet: Rabbit polyclonal anti-STX7 , Bethyl , Cat#A304-512A; RRID: AB_2620706.



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    Image Search Results


    Figure 4. DEP impair autophagic flux in the degradation stage. (a) HUVECs infected with RFP-LC3B-GFP- expressing adenovirus were exposed to DEP (70 µg/ml) for 18 h and autolysosome formation was examined under fluorescence microscopy. (b) The number of yellow signals per cell was counted from 10 randomly selected cells. Statistical analysis was performed using two-tailed Student’s t test. **P < 0.02 versus control. (c) HUVECs were pre-incubated with Bafilomysin A1 (Baf A1, 100 nM) for 3 h and exposed to DEP (70 μg/ ml) for an additional 12 h. LC3B level was measured by immunoblotting. (d) Quantification of LC3BII levels normalized to β-actin. Results are presented as means ± SD (n = 3). Statistical analysis was performed using one-way ANOVA. **P < 0.02 versus no treatment. (e) HUVECs were exposed to DEP (70 μg/ml) for 24 h and protein levels of STX17, VAMP8, SNAP29, YKT6, STX7, and β-actin were measured by immunoblotting. (f) Quantification of STX17, VAMP8, SNAP29, YKT6, and STX7 levels normalized to β-actin. Results are presented as means ± SD (n = 3). Statistical analysis was performed using two-tailed Student’s t test. **P < 0.02 versus control.

    Journal: Scientific reports

    Article Title: Diesel exhaust particles induce human umbilical vein endothelial cells apoptosis by accumulation of autophagosomes and caspase-8 activation.

    doi: 10.1038/s41598-022-21044-3

    Figure Lengend Snippet: Figure 4. DEP impair autophagic flux in the degradation stage. (a) HUVECs infected with RFP-LC3B-GFP- expressing adenovirus were exposed to DEP (70 µg/ml) for 18 h and autolysosome formation was examined under fluorescence microscopy. (b) The number of yellow signals per cell was counted from 10 randomly selected cells. Statistical analysis was performed using two-tailed Student’s t test. **P < 0.02 versus control. (c) HUVECs were pre-incubated with Bafilomysin A1 (Baf A1, 100 nM) for 3 h and exposed to DEP (70 μg/ ml) for an additional 12 h. LC3B level was measured by immunoblotting. (d) Quantification of LC3BII levels normalized to β-actin. Results are presented as means ± SD (n = 3). Statistical analysis was performed using one-way ANOVA. **P < 0.02 versus no treatment. (e) HUVECs were exposed to DEP (70 μg/ml) for 24 h and protein levels of STX17, VAMP8, SNAP29, YKT6, STX7, and β-actin were measured by immunoblotting. (f) Quantification of STX17, VAMP8, SNAP29, YKT6, and STX7 levels normalized to β-actin. Results are presented as means ± SD (n = 3). Statistical analysis was performed using two-tailed Student’s t test. **P < 0.02 versus control.

    Article Snippet: The STX7 antibody was purchased from Bethyl Laboratories Inc. (Montgomery, TX, USA).

    Techniques: Infection, Expressing, Fluorescence, Microscopy, Two Tailed Test, Control, Incubation, Western Blot

    DEP impair autophagic flux in the degradation stage. ( a ) HUVECs infected with RFP-LC3B-GFP-expressing adenovirus were exposed to DEP (70 µg/ml) for 18 h and autolysosome formation was examined under fluorescence microscopy. ( b ) The number of yellow signals per cell was counted from 10 randomly selected cells. Statistical analysis was performed using two-tailed Student’s t test. **P < 0.02 versus control. ( c ) HUVECs were pre-incubated with Bafilomysin A1 (Baf A1, 100 nM) for 3 h and exposed to DEP (70 μg/ml) for an additional 12 h. LC3B level was measured by immunoblotting. ( d ) Quantification of LC3BII levels normalized to β-actin. Results are presented as means ± SD ( n = 3). Statistical analysis was performed using one-way ANOVA. **P < 0.02 versus no treatment. ( e ) HUVECs were exposed to DEP (70 μg/ml) for 24 h and protein levels of STX17, VAMP8, SNAP29, YKT6, STX7, and β-actin were measured by immunoblotting. ( f ) Quantification of STX17, VAMP8, SNAP29, YKT6, and STX7 levels normalized to β-actin. Results are presented as means ± SD ( n = 3). Statistical analysis was performed using two-tailed Student’s t test. **P < 0.02 versus control.

    Journal: Scientific Reports

    Article Title: Diesel exhaust particles induce human umbilical vein endothelial cells apoptosis by accumulation of autophagosomes and caspase-8 activation

    doi: 10.1038/s41598-022-21044-3

    Figure Lengend Snippet: DEP impair autophagic flux in the degradation stage. ( a ) HUVECs infected with RFP-LC3B-GFP-expressing adenovirus were exposed to DEP (70 µg/ml) for 18 h and autolysosome formation was examined under fluorescence microscopy. ( b ) The number of yellow signals per cell was counted from 10 randomly selected cells. Statistical analysis was performed using two-tailed Student’s t test. **P < 0.02 versus control. ( c ) HUVECs were pre-incubated with Bafilomysin A1 (Baf A1, 100 nM) for 3 h and exposed to DEP (70 μg/ml) for an additional 12 h. LC3B level was measured by immunoblotting. ( d ) Quantification of LC3BII levels normalized to β-actin. Results are presented as means ± SD ( n = 3). Statistical analysis was performed using one-way ANOVA. **P < 0.02 versus no treatment. ( e ) HUVECs were exposed to DEP (70 μg/ml) for 24 h and protein levels of STX17, VAMP8, SNAP29, YKT6, STX7, and β-actin were measured by immunoblotting. ( f ) Quantification of STX17, VAMP8, SNAP29, YKT6, and STX7 levels normalized to β-actin. Results are presented as means ± SD ( n = 3). Statistical analysis was performed using two-tailed Student’s t test. **P < 0.02 versus control.

    Article Snippet: The STX7 antibody was purchased from Bethyl Laboratories Inc. (Montgomery, TX, USA).

    Techniques: Infection, Expressing, Fluorescence, Microscopy, Two Tailed Test, Incubation, Western Blot

    Journal: iScience

    Article Title: Persistent coxsackievirus B1 infection triggers extensive changes in the transcriptome of human pancreatic ductal cells

    doi: 10.1016/j.isci.2021.103653

    Figure Lengend Snippet:

    Article Snippet: Rabbit polyclonal anti-STX7 , Bethyl , Cat#A304-512A; RRID: AB_2620706.

    Techniques: Immunofluorescence, Virus, Control, Recombinant, Reverse Transcription, Gene Expression, Software, Transformation Assay