anti atg5 ab Search Results


anti atg5 antibody  (Novus Biologicals)
94
Novus Biologicals anti atg5 antibody
TRIM44 promtoes aggregates deaggregation and clearance via autophagy. (a) Cells were treated with MG132 and immunostained with antibodies to ubiquitin (red) and LC3B-II (green). Arrows indicate ubiquitin-positive aggregates that colocalize with LC3B-positive autophagosomes. Scale bars: 10 μm. (b) Confocal images of TRIM44[OE-CON] and TRIM44[OE] U266 cells after treatment with MG132 (0.5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA. Arrows indicate cells with remaining aggregates. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA (10 mM) and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (0.5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. (c) Confocal images of TRIM44[KD-CON] and TRIM44[KD] U266 cells after treatment with MG132 (0.5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or PP242 (10 nM). Arrows indicate cells with remaining aggregates. Scale bars: 10 µm. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or PP242 (10 nM) and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (0.5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. (d, e) Confocal images of WT or <t>ATG5</t> KO TRIM44[OE-CON] and TRIM44[OE] U266 cells transfected with NC or ATG5 siRNA after treatment with MG132 (0.5 µM) for 16 h. Aggregates (marked by arrows) were identified by staining with the antibody against ubiquitin (d). Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of ATG5 and TRIM44 were assayed by western blots (e). (f, g) Confocal images of WT or BECN1 KO TRIM44[OE-CON] and TRIM44[OE] U266 cells were treated with MG132 (0.5 µM) for 16 h. Aggregates (marked by arrows) were identified by staining with the antibody against ubiquitin (f). Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of BECN1 and TRIM44 were assayed by western blot (g). (h, i) Confocal images of RPMI-TRIM44[Tet-on] cells treated with or without DOX (1 µg/mL) transfected with NC or ATG5 siRNA after treatment with MG132 (0.5 µM) for 16 h. The status of aggregates after MG132 treatment was quantified in the histogram. Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of ATG5 and TRIM44 were assayed by western blots (h). (j) Confocal images of RPMI-TRIM44[Tet-on] cells treated with or without DOX (1 µg/mL) together with MG132 (5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA. Arrows indicate cells with remaining aggregates. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm.
Anti Atg5 Antibody, supplied by Novus Biologicals, 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 atg5 antibody/product/Novus Biologicals
Average 94 stars, based on 1 article reviews
anti atg5 antibody - by Bioz Stars, 2026-03
94/100 stars
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anti atg5  (Cell Signaling Technology Inc)
97
Cell Signaling Technology Inc anti atg5
TRIM44 promtoes aggregates deaggregation and clearance via autophagy. (a) Cells were treated with MG132 and immunostained with antibodies to ubiquitin (red) and LC3B-II (green). Arrows indicate ubiquitin-positive aggregates that colocalize with LC3B-positive autophagosomes. Scale bars: 10 μm. (b) Confocal images of TRIM44[OE-CON] and TRIM44[OE] U266 cells after treatment with MG132 (0.5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA. Arrows indicate cells with remaining aggregates. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA (10 mM) and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (0.5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. (c) Confocal images of TRIM44[KD-CON] and TRIM44[KD] U266 cells after treatment with MG132 (0.5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or PP242 (10 nM). Arrows indicate cells with remaining aggregates. Scale bars: 10 µm. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or PP242 (10 nM) and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (0.5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. (d, e) Confocal images of WT or <t>ATG5</t> KO TRIM44[OE-CON] and TRIM44[OE] U266 cells transfected with NC or ATG5 siRNA after treatment with MG132 (0.5 µM) for 16 h. Aggregates (marked by arrows) were identified by staining with the antibody against ubiquitin (d). Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of ATG5 and TRIM44 were assayed by western blots (e). (f, g) Confocal images of WT or BECN1 KO TRIM44[OE-CON] and TRIM44[OE] U266 cells were treated with MG132 (0.5 µM) for 16 h. Aggregates (marked by arrows) were identified by staining with the antibody against ubiquitin (f). Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of BECN1 and TRIM44 were assayed by western blot (g). (h, i) Confocal images of RPMI-TRIM44[Tet-on] cells treated with or without DOX (1 µg/mL) transfected with NC or ATG5 siRNA after treatment with MG132 (0.5 µM) for 16 h. The status of aggregates after MG132 treatment was quantified in the histogram. Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of ATG5 and TRIM44 were assayed by western blots (h). (j) Confocal images of RPMI-TRIM44[Tet-on] cells treated with or without DOX (1 µg/mL) together with MG132 (5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA. Arrows indicate cells with remaining aggregates. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm.
Anti Atg5, supplied by Cell Signaling Technology Inc, 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/anti atg5/product/Cell Signaling Technology Inc
Average 97 stars, based on 1 article reviews
anti atg5 - by Bioz Stars, 2026-03
97/100 stars
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anti atg5  (Santa Cruz Biotechnology)
96
Santa Cruz Biotechnology anti atg5
TRIM44 promtoes aggregates deaggregation and clearance via autophagy. (a) Cells were treated with MG132 and immunostained with antibodies to ubiquitin (red) and LC3B-II (green). Arrows indicate ubiquitin-positive aggregates that colocalize with LC3B-positive autophagosomes. Scale bars: 10 μm. (b) Confocal images of TRIM44[OE-CON] and TRIM44[OE] U266 cells after treatment with MG132 (0.5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA. Arrows indicate cells with remaining aggregates. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA (10 mM) and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (0.5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. (c) Confocal images of TRIM44[KD-CON] and TRIM44[KD] U266 cells after treatment with MG132 (0.5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or PP242 (10 nM). Arrows indicate cells with remaining aggregates. Scale bars: 10 µm. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or PP242 (10 nM) and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (0.5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. (d, e) Confocal images of WT or <t>ATG5</t> KO TRIM44[OE-CON] and TRIM44[OE] U266 cells transfected with NC or ATG5 siRNA after treatment with MG132 (0.5 µM) for 16 h. Aggregates (marked by arrows) were identified by staining with the antibody against ubiquitin (d). Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of ATG5 and TRIM44 were assayed by western blots (e). (f, g) Confocal images of WT or BECN1 KO TRIM44[OE-CON] and TRIM44[OE] U266 cells were treated with MG132 (0.5 µM) for 16 h. Aggregates (marked by arrows) were identified by staining with the antibody against ubiquitin (f). Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of BECN1 and TRIM44 were assayed by western blot (g). (h, i) Confocal images of RPMI-TRIM44[Tet-on] cells treated with or without DOX (1 µg/mL) transfected with NC or ATG5 siRNA after treatment with MG132 (0.5 µM) for 16 h. The status of aggregates after MG132 treatment was quantified in the histogram. Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of ATG5 and TRIM44 were assayed by western blots (h). (j) Confocal images of RPMI-TRIM44[Tet-on] cells treated with or without DOX (1 µg/mL) together with MG132 (5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA. Arrows indicate cells with remaining aggregates. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm.
Anti Atg5, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti atg5/product/Santa Cruz Biotechnology
Average 96 stars, based on 1 article reviews
anti atg5 - by Bioz Stars, 2026-03
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anti atg5 mab  (Cell Signaling Technology Inc)
97
Cell Signaling Technology Inc anti atg5 mab
TRIM44 promtoes aggregates deaggregation and clearance via autophagy. (a) Cells were treated with MG132 and immunostained with antibodies to ubiquitin (red) and LC3B-II (green). Arrows indicate ubiquitin-positive aggregates that colocalize with LC3B-positive autophagosomes. Scale bars: 10 μm. (b) Confocal images of TRIM44[OE-CON] and TRIM44[OE] U266 cells after treatment with MG132 (0.5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA. Arrows indicate cells with remaining aggregates. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA (10 mM) and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (0.5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. (c) Confocal images of TRIM44[KD-CON] and TRIM44[KD] U266 cells after treatment with MG132 (0.5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or PP242 (10 nM). Arrows indicate cells with remaining aggregates. Scale bars: 10 µm. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or PP242 (10 nM) and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (0.5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. (d, e) Confocal images of WT or <t>ATG5</t> KO TRIM44[OE-CON] and TRIM44[OE] U266 cells transfected with NC or ATG5 siRNA after treatment with MG132 (0.5 µM) for 16 h. Aggregates (marked by arrows) were identified by staining with the antibody against ubiquitin (d). Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of ATG5 and TRIM44 were assayed by western blots (e). (f, g) Confocal images of WT or BECN1 KO TRIM44[OE-CON] and TRIM44[OE] U266 cells were treated with MG132 (0.5 µM) for 16 h. Aggregates (marked by arrows) were identified by staining with the antibody against ubiquitin (f). Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of BECN1 and TRIM44 were assayed by western blot (g). (h, i) Confocal images of RPMI-TRIM44[Tet-on] cells treated with or without DOX (1 µg/mL) transfected with NC or ATG5 siRNA after treatment with MG132 (0.5 µM) for 16 h. The status of aggregates after MG132 treatment was quantified in the histogram. Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of ATG5 and TRIM44 were assayed by western blots (h). (j) Confocal images of RPMI-TRIM44[Tet-on] cells treated with or without DOX (1 µg/mL) together with MG132 (5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA. Arrows indicate cells with remaining aggregates. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm.
Anti Atg5 Mab, supplied by Cell Signaling Technology Inc, 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/anti atg5 mab/product/Cell Signaling Technology Inc
Average 97 stars, based on 1 article reviews
anti atg5 mab - by Bioz Stars, 2026-03
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atg5  (Novus Biologicals)
95
Novus Biologicals atg5
The role of autophagy in morphine-mediated synaptic alterations. Representative Western blot (A) and quantification of autophagy mediators (Beclin1, <t>ATG5,</t> p62, and LC3-II; B) in the hippocampal lysates from mouse injected with saline or morphine ( n = 3 to 4/group). *, P < 0.05; **, P < 0.01 versus saline group using Student’s t test. Representative time course Western blot (C) and quantification of LC3-II (D) in primary neurons exposed to morphine. Representative Western blot (E) and quantification of Beclin1, ATG5, p62, and LC3-II (F) in the cell lysates from neurons pretreated with naltrexone for 1 h followed by 24-h morphine treatment. Representative confocal images (G) and quantification of GFP-LC3 puncta (H) of primary hippocampal neurons expressing GFP-LC3 exposed to morphine for 24 h. Bar, 10 µm. **, P < 0.01 versus saline group using Student's t test. Representative Western blot (I) and quantification of LC3-II (J) in the cell lysates of neurons treated with bafilomycin for 4 h after 24-h exposure to morphine. Representative confocal images of GFP-expressing primary rat hippocampal neurons treated with wortmannin (500 µM) for 1 h followed by morphine exposure for 24 h and stained with vGlut1 and GAD65 (K) and quantifications of spine density and excitatory and inhibitory synapses (L). Bar, 5 µm. Representative confocal images of neurons expressing PGK-GFP-scramble or PGK-GFP-shATG7 vector in the presence or absence of morphine for 24 h and stained with vGlut1 and GAD65 (M) and quantifications of spine density and excitatory and inhibitory synapses (N). Bar, 5 µm. Quantification of Western blot results were all normalized to β-actin. Each set of in vitro results was quantified upon four independent experiments. All data are presented as mean ± SD. *, P < 0.05; **, P < 0.01 versus control group; # , P < 0.05; ## , P < 0.01 versus morphine group using one-way ANOVA with post hoc test (except B and H).
Atg5, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/atg5/product/Novus Biologicals
Average 95 stars, based on 1 article reviews
atg5 - by Bioz Stars, 2026-03
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anti-atg5  (Thermo Fisher)
90
Thermo Fisher anti-atg5
Exposure to a mixture of 7-oxysterols (2mix) causes lipid accumulation and autophagy dysfunction. THP-1 monocytes or THP-1 differentiated macrophages were treated with or without 2mix for 12 hours to 48 hours. In some experiments, cells were pretreated with the autophagy inhibitor 3MA or the autophagy inducer rapamycin for 1 hour and then exposed to 2mix or treated with the lysosomatropic agent CQ for 24 hours or CQ for 1 hour and 2mix for 24 hours. ( A ) Oil red O-stained THP-1 monocytes counterstained with hematoxylin (24 hours), bars = 100 µm. ( B and C ) Accumulation of autophagy vacuoles in THP-1 differentiated macrophages assayed by TEM and quantified by image analysis. ( B ) Photographs of low power view of control and 2mix-treated cells (two photographs on the left-hand side), bars = 2 µm or a higher magnified photograph (photograph on the top right-hand side) demonstrates autophagic vacuoles with double-membrane structure containing undigested cytoplasmic materials, bar = 1 µm. ( C ) Quantification of numbers of autophagic vacuoles, * P < 0.05, ** P < 0.01 versus control and 3MA + 2mix, respectively. ( D ) <t>Atg5-immunostained</t> THP-1 monocytes analyzed by flow cytometry. n = 6–8, * P < 0.05, ** P < 0.01 and *** P < 0.001. The inset shows a representative histogram of flow cytometry results (control: gray field histogram; 2mix: black-lined empty histogram). ( E ) LC3β-immunostained THP-1 monocytes analyzed by flow cytometry; n = 6–20, * P < 0.05. ( F ) Western blot analysis of LC3 I/II in the whole cell lysate of THP-1 monocytes. Values mentioned below the Western blot image are the ratios of LC3-II to LC3-I. ( G ) p62/ SQSTM1 -immunostained THP-1 monocytes analyzed by flow cytometry; n = 3–4. a P < 0.05 vs CQ and P < 0.01 vs 3MA + 2mix and CQ + 2mix. b P < 0.05 vs CQ and P < 0.01 vs CQ + 2mix and P < 0.001 vs 3MA + 2mix. * P < 0.05.
Anti Atg5, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti-atg5/product/Thermo Fisher
Average 90 stars, based on 1 article reviews
anti-atg5 - by Bioz Stars, 2026-03
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anti-atg5  (Millipore)
90
Millipore anti-atg5
Exposure to a mixture of 7-oxysterols (2mix) causes lipid accumulation and autophagy dysfunction. THP-1 monocytes or THP-1 differentiated macrophages were treated with or without 2mix for 12 hours to 48 hours. In some experiments, cells were pretreated with the autophagy inhibitor 3MA or the autophagy inducer rapamycin for 1 hour and then exposed to 2mix or treated with the lysosomatropic agent CQ for 24 hours or CQ for 1 hour and 2mix for 24 hours. ( A ) Oil red O-stained THP-1 monocytes counterstained with hematoxylin (24 hours), bars = 100 µm. ( B and C ) Accumulation of autophagy vacuoles in THP-1 differentiated macrophages assayed by TEM and quantified by image analysis. ( B ) Photographs of low power view of control and 2mix-treated cells (two photographs on the left-hand side), bars = 2 µm or a higher magnified photograph (photograph on the top right-hand side) demonstrates autophagic vacuoles with double-membrane structure containing undigested cytoplasmic materials, bar = 1 µm. ( C ) Quantification of numbers of autophagic vacuoles, * P < 0.05, ** P < 0.01 versus control and 3MA + 2mix, respectively. ( D ) <t>Atg5-immunostained</t> THP-1 monocytes analyzed by flow cytometry. n = 6–8, * P < 0.05, ** P < 0.01 and *** P < 0.001. The inset shows a representative histogram of flow cytometry results (control: gray field histogram; 2mix: black-lined empty histogram). ( E ) LC3β-immunostained THP-1 monocytes analyzed by flow cytometry; n = 6–20, * P < 0.05. ( F ) Western blot analysis of LC3 I/II in the whole cell lysate of THP-1 monocytes. Values mentioned below the Western blot image are the ratios of LC3-II to LC3-I. ( G ) p62/ SQSTM1 -immunostained THP-1 monocytes analyzed by flow cytometry; n = 3–4. a P < 0.05 vs CQ and P < 0.01 vs 3MA + 2mix and CQ + 2mix. b P < 0.05 vs CQ and P < 0.01 vs CQ + 2mix and P < 0.001 vs 3MA + 2mix. * P < 0.05.
Anti Atg5, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti-atg5 - by Bioz Stars, 2026-03
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antibodies against atg5  (Beyotime)
90
Beyotime antibodies against atg5
Exposure to a mixture of 7-oxysterols (2mix) causes lipid accumulation and autophagy dysfunction. THP-1 monocytes or THP-1 differentiated macrophages were treated with or without 2mix for 12 hours to 48 hours. In some experiments, cells were pretreated with the autophagy inhibitor 3MA or the autophagy inducer rapamycin for 1 hour and then exposed to 2mix or treated with the lysosomatropic agent CQ for 24 hours or CQ for 1 hour and 2mix for 24 hours. ( A ) Oil red O-stained THP-1 monocytes counterstained with hematoxylin (24 hours), bars = 100 µm. ( B and C ) Accumulation of autophagy vacuoles in THP-1 differentiated macrophages assayed by TEM and quantified by image analysis. ( B ) Photographs of low power view of control and 2mix-treated cells (two photographs on the left-hand side), bars = 2 µm or a higher magnified photograph (photograph on the top right-hand side) demonstrates autophagic vacuoles with double-membrane structure containing undigested cytoplasmic materials, bar = 1 µm. ( C ) Quantification of numbers of autophagic vacuoles, * P < 0.05, ** P < 0.01 versus control and 3MA + 2mix, respectively. ( D ) <t>Atg5-immunostained</t> THP-1 monocytes analyzed by flow cytometry. n = 6–8, * P < 0.05, ** P < 0.01 and *** P < 0.001. The inset shows a representative histogram of flow cytometry results (control: gray field histogram; 2mix: black-lined empty histogram). ( E ) LC3β-immunostained THP-1 monocytes analyzed by flow cytometry; n = 6–20, * P < 0.05. ( F ) Western blot analysis of LC3 I/II in the whole cell lysate of THP-1 monocytes. Values mentioned below the Western blot image are the ratios of LC3-II to LC3-I. ( G ) p62/ SQSTM1 -immunostained THP-1 monocytes analyzed by flow cytometry; n = 3–4. a P < 0.05 vs CQ and P < 0.01 vs 3MA + 2mix and CQ + 2mix. b P < 0.05 vs CQ and P < 0.01 vs CQ + 2mix and P < 0.001 vs 3MA + 2mix. * P < 0.05.
Antibodies Against Atg5, supplied by Beyotime, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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antibodies against atg5 - by Bioz Stars, 2026-03
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anti atg5  (Proteintech)
96
Proteintech anti atg5
Exposure to a mixture of 7-oxysterols (2mix) causes lipid accumulation and autophagy dysfunction. THP-1 monocytes or THP-1 differentiated macrophages were treated with or without 2mix for 12 hours to 48 hours. In some experiments, cells were pretreated with the autophagy inhibitor 3MA or the autophagy inducer rapamycin for 1 hour and then exposed to 2mix or treated with the lysosomatropic agent CQ for 24 hours or CQ for 1 hour and 2mix for 24 hours. ( A ) Oil red O-stained THP-1 monocytes counterstained with hematoxylin (24 hours), bars = 100 µm. ( B and C ) Accumulation of autophagy vacuoles in THP-1 differentiated macrophages assayed by TEM and quantified by image analysis. ( B ) Photographs of low power view of control and 2mix-treated cells (two photographs on the left-hand side), bars = 2 µm or a higher magnified photograph (photograph on the top right-hand side) demonstrates autophagic vacuoles with double-membrane structure containing undigested cytoplasmic materials, bar = 1 µm. ( C ) Quantification of numbers of autophagic vacuoles, * P < 0.05, ** P < 0.01 versus control and 3MA + 2mix, respectively. ( D ) <t>Atg5-immunostained</t> THP-1 monocytes analyzed by flow cytometry. n = 6–8, * P < 0.05, ** P < 0.01 and *** P < 0.001. The inset shows a representative histogram of flow cytometry results (control: gray field histogram; 2mix: black-lined empty histogram). ( E ) LC3β-immunostained THP-1 monocytes analyzed by flow cytometry; n = 6–20, * P < 0.05. ( F ) Western blot analysis of LC3 I/II in the whole cell lysate of THP-1 monocytes. Values mentioned below the Western blot image are the ratios of LC3-II to LC3-I. ( G ) p62/ SQSTM1 -immunostained THP-1 monocytes analyzed by flow cytometry; n = 3–4. a P < 0.05 vs CQ and P < 0.01 vs 3MA + 2mix and CQ + 2mix. b P < 0.05 vs CQ and P < 0.01 vs CQ + 2mix and P < 0.001 vs 3MA + 2mix. * P < 0.05.
Anti Atg5, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti atg5/product/Proteintech
Average 96 stars, based on 1 article reviews
anti atg5 - by Bioz Stars, 2026-03
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anti-atg5  (Cosmo Bio USA)
90
Cosmo Bio USA anti-atg5
Exposure to a mixture of 7-oxysterols (2mix) causes lipid accumulation and autophagy dysfunction. THP-1 monocytes or THP-1 differentiated macrophages were treated with or without 2mix for 12 hours to 48 hours. In some experiments, cells were pretreated with the autophagy inhibitor 3MA or the autophagy inducer rapamycin for 1 hour and then exposed to 2mix or treated with the lysosomatropic agent CQ for 24 hours or CQ for 1 hour and 2mix for 24 hours. ( A ) Oil red O-stained THP-1 monocytes counterstained with hematoxylin (24 hours), bars = 100 µm. ( B and C ) Accumulation of autophagy vacuoles in THP-1 differentiated macrophages assayed by TEM and quantified by image analysis. ( B ) Photographs of low power view of control and 2mix-treated cells (two photographs on the left-hand side), bars = 2 µm or a higher magnified photograph (photograph on the top right-hand side) demonstrates autophagic vacuoles with double-membrane structure containing undigested cytoplasmic materials, bar = 1 µm. ( C ) Quantification of numbers of autophagic vacuoles, * P < 0.05, ** P < 0.01 versus control and 3MA + 2mix, respectively. ( D ) <t>Atg5-immunostained</t> THP-1 monocytes analyzed by flow cytometry. n = 6–8, * P < 0.05, ** P < 0.01 and *** P < 0.001. The inset shows a representative histogram of flow cytometry results (control: gray field histogram; 2mix: black-lined empty histogram). ( E ) LC3β-immunostained THP-1 monocytes analyzed by flow cytometry; n = 6–20, * P < 0.05. ( F ) Western blot analysis of LC3 I/II in the whole cell lysate of THP-1 monocytes. Values mentioned below the Western blot image are the ratios of LC3-II to LC3-I. ( G ) p62/ SQSTM1 -immunostained THP-1 monocytes analyzed by flow cytometry; n = 3–4. a P < 0.05 vs CQ and P < 0.01 vs 3MA + 2mix and CQ + 2mix. b P < 0.05 vs CQ and P < 0.01 vs CQ + 2mix and P < 0.001 vs 3MA + 2mix. * P < 0.05.
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TRIM44 promtoes aggregates deaggregation and clearance via autophagy. (a) Cells were treated with MG132 and immunostained with antibodies to ubiquitin (red) and LC3B-II (green). Arrows indicate ubiquitin-positive aggregates that colocalize with LC3B-positive autophagosomes. Scale bars: 10 μm. (b) Confocal images of TRIM44[OE-CON] and TRIM44[OE] U266 cells after treatment with MG132 (0.5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA. Arrows indicate cells with remaining aggregates. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA (10 mM) and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (0.5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. (c) Confocal images of TRIM44[KD-CON] and TRIM44[KD] U266 cells after treatment with MG132 (0.5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or PP242 (10 nM). Arrows indicate cells with remaining aggregates. Scale bars: 10 µm. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or PP242 (10 nM) and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (0.5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. (d, e) Confocal images of WT or ATG5 KO TRIM44[OE-CON] and TRIM44[OE] U266 cells transfected with NC or ATG5 siRNA after treatment with MG132 (0.5 µM) for 16 h. Aggregates (marked by arrows) were identified by staining with the antibody against ubiquitin (d). Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of ATG5 and TRIM44 were assayed by western blots (e). (f, g) Confocal images of WT or BECN1 KO TRIM44[OE-CON] and TRIM44[OE] U266 cells were treated with MG132 (0.5 µM) for 16 h. Aggregates (marked by arrows) were identified by staining with the antibody against ubiquitin (f). Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of BECN1 and TRIM44 were assayed by western blot (g). (h, i) Confocal images of RPMI-TRIM44[Tet-on] cells treated with or without DOX (1 µg/mL) transfected with NC or ATG5 siRNA after treatment with MG132 (0.5 µM) for 16 h. The status of aggregates after MG132 treatment was quantified in the histogram. Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of ATG5 and TRIM44 were assayed by western blots (h). (j) Confocal images of RPMI-TRIM44[Tet-on] cells treated with or without DOX (1 µg/mL) together with MG132 (5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA. Arrows indicate cells with remaining aggregates. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm.

Journal: Autophagy

Article Title: TRIM44 links the UPS to SQSTM1/p62-dependent aggrephagy and removing misfolded proteins

doi: 10.1080/15548627.2021.1956105

Figure Lengend Snippet: TRIM44 promtoes aggregates deaggregation and clearance via autophagy. (a) Cells were treated with MG132 and immunostained with antibodies to ubiquitin (red) and LC3B-II (green). Arrows indicate ubiquitin-positive aggregates that colocalize with LC3B-positive autophagosomes. Scale bars: 10 μm. (b) Confocal images of TRIM44[OE-CON] and TRIM44[OE] U266 cells after treatment with MG132 (0.5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA. Arrows indicate cells with remaining aggregates. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA (10 mM) and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (0.5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. (c) Confocal images of TRIM44[KD-CON] and TRIM44[KD] U266 cells after treatment with MG132 (0.5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or PP242 (10 nM). Arrows indicate cells with remaining aggregates. Scale bars: 10 µm. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or PP242 (10 nM) and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (0.5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. (d, e) Confocal images of WT or ATG5 KO TRIM44[OE-CON] and TRIM44[OE] U266 cells transfected with NC or ATG5 siRNA after treatment with MG132 (0.5 µM) for 16 h. Aggregates (marked by arrows) were identified by staining with the antibody against ubiquitin (d). Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of ATG5 and TRIM44 were assayed by western blots (e). (f, g) Confocal images of WT or BECN1 KO TRIM44[OE-CON] and TRIM44[OE] U266 cells were treated with MG132 (0.5 µM) for 16 h. Aggregates (marked by arrows) were identified by staining with the antibody against ubiquitin (f). Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of BECN1 and TRIM44 were assayed by western blot (g). (h, i) Confocal images of RPMI-TRIM44[Tet-on] cells treated with or without DOX (1 µg/mL) transfected with NC or ATG5 siRNA after treatment with MG132 (0.5 µM) for 16 h. The status of aggregates after MG132 treatment was quantified in the histogram. Scale bars: 10 µm. The status of aggregates after MG132 treatment was quantified in the histogram. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm. The protein level of ATG5 and TRIM44 were assayed by western blots (h). (j) Confocal images of RPMI-TRIM44[Tet-on] cells treated with or without DOX (1 µg/mL) together with MG132 (5 µM) for 16 h followed by a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA. Arrows indicate cells with remaining aggregates. The relative level of remaining aggregates is determinded by quantifying the percentage of cells with remaining aggregates after a 24-h chase period in normal culture media with DMSO (vehicle), or 3-MA and normalized to the percentage of cells with aggregates formed by the 16 h MG132 (5 µM) treatment in corresponding cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Scale bars: 10 μm.

Article Snippet: Anti-TRIM44 polyclonal antibody (Proteintech Group, 11,511-1-AP); anti-Ub antibody (Biolegend, 646,301); anti-mCherry antibody (ThermoFisher, PA5-34,974); anti-VIM antibody (ThermoFisher, MA5-11,883); anti-NFE2L2/NRF2 antibody (ThermoFisher, PA5-27,882); anti-20S proteasome antibody (MilliporeSigma, ST1049); anti-TUBG/γ-Tubulin antibody (MilliporeSigma, T5326); anti-ATG5 antibody (Novus Biologicals, NBP2-24,389); Anti-GFP antibody (Santa Cruz Biotechnology, sc-9996); anti-ACTB/β-Actin antibody (Santa Cruz Biotechnology, sc-47,778), anti-HA antibody (Santa Cruz Biotechnology, sc-805); anti-BECN1 antibody (Cell Signaling Technology, 4122); anti-HDAC6 antibody (Cell Signaling Technology, 7558); anti-LC3B antibody (Cell Signaling Technology, 3868); anti-SQSTM1/p62 antibody (Cell Signaling Technology, 88,588); anti-cleaved PARP antibody (Cell Signaling Technology, 5625); Alexa Fluor 594 donkey anti rabbit IgG(H + L) (Invitrogen, A21207); Alexa Fluor 546 goat anti mouse IgG(H + L) (Invitrogen, A11003).

Techniques: Ubiquitin Proteomics, Transfection, Staining, Western Blot

The role of autophagy in morphine-mediated synaptic alterations. Representative Western blot (A) and quantification of autophagy mediators (Beclin1, ATG5, p62, and LC3-II; B) in the hippocampal lysates from mouse injected with saline or morphine ( n = 3 to 4/group). *, P < 0.05; **, P < 0.01 versus saline group using Student’s t test. Representative time course Western blot (C) and quantification of LC3-II (D) in primary neurons exposed to morphine. Representative Western blot (E) and quantification of Beclin1, ATG5, p62, and LC3-II (F) in the cell lysates from neurons pretreated with naltrexone for 1 h followed by 24-h morphine treatment. Representative confocal images (G) and quantification of GFP-LC3 puncta (H) of primary hippocampal neurons expressing GFP-LC3 exposed to morphine for 24 h. Bar, 10 µm. **, P < 0.01 versus saline group using Student's t test. Representative Western blot (I) and quantification of LC3-II (J) in the cell lysates of neurons treated with bafilomycin for 4 h after 24-h exposure to morphine. Representative confocal images of GFP-expressing primary rat hippocampal neurons treated with wortmannin (500 µM) for 1 h followed by morphine exposure for 24 h and stained with vGlut1 and GAD65 (K) and quantifications of spine density and excitatory and inhibitory synapses (L). Bar, 5 µm. Representative confocal images of neurons expressing PGK-GFP-scramble or PGK-GFP-shATG7 vector in the presence or absence of morphine for 24 h and stained with vGlut1 and GAD65 (M) and quantifications of spine density and excitatory and inhibitory synapses (N). Bar, 5 µm. Quantification of Western blot results were all normalized to β-actin. Each set of in vitro results was quantified upon four independent experiments. All data are presented as mean ± SD. *, P < 0.05; **, P < 0.01 versus control group; # , P < 0.05; ## , P < 0.01 versus morphine group using one-way ANOVA with post hoc test (except B and H).

Journal: The Journal of Cell Biology

Article Title: Regulation of morphine-induced synaptic alterations: Role of oxidative stress, ER stress, and autophagy

doi: 10.1083/jcb.201605065

Figure Lengend Snippet: The role of autophagy in morphine-mediated synaptic alterations. Representative Western blot (A) and quantification of autophagy mediators (Beclin1, ATG5, p62, and LC3-II; B) in the hippocampal lysates from mouse injected with saline or morphine ( n = 3 to 4/group). *, P < 0.05; **, P < 0.01 versus saline group using Student’s t test. Representative time course Western blot (C) and quantification of LC3-II (D) in primary neurons exposed to morphine. Representative Western blot (E) and quantification of Beclin1, ATG5, p62, and LC3-II (F) in the cell lysates from neurons pretreated with naltrexone for 1 h followed by 24-h morphine treatment. Representative confocal images (G) and quantification of GFP-LC3 puncta (H) of primary hippocampal neurons expressing GFP-LC3 exposed to morphine for 24 h. Bar, 10 µm. **, P < 0.01 versus saline group using Student's t test. Representative Western blot (I) and quantification of LC3-II (J) in the cell lysates of neurons treated with bafilomycin for 4 h after 24-h exposure to morphine. Representative confocal images of GFP-expressing primary rat hippocampal neurons treated with wortmannin (500 µM) for 1 h followed by morphine exposure for 24 h and stained with vGlut1 and GAD65 (K) and quantifications of spine density and excitatory and inhibitory synapses (L). Bar, 5 µm. Representative confocal images of neurons expressing PGK-GFP-scramble or PGK-GFP-shATG7 vector in the presence or absence of morphine for 24 h and stained with vGlut1 and GAD65 (M) and quantifications of spine density and excitatory and inhibitory synapses (N). Bar, 5 µm. Quantification of Western blot results were all normalized to β-actin. Each set of in vitro results was quantified upon four independent experiments. All data are presented as mean ± SD. *, P < 0.05; **, P < 0.01 versus control group; # , P < 0.05; ## , P < 0.01 versus morphine group using one-way ANOVA with post hoc test (except B and H).

Article Snippet: The Western blots were then probed with respective antibodies recognizing the vGlut1 (1:5,000; catalog no. AB5905, lot no. RRID:AB_2301751; EMD Millipore), PSD95 (1:1,000; catalog no. ab18258; lot no. RRID:AB_444362; Abcam), GAD65 (1:1,000; catalog no. NBP1-33284, lot no. RRID:AB_10004060; Novus Biologicals), gephyrin (1:250; catalog no. 610584, lot no. RRID:AB_397929; BD), BIP (1:1,000; catalog no. 610978, lot no. RRID:AB_398291; BD), pPERK (1:250; catalog no. sc-32577, lot no. RRID:AB_2293243; Santa Cruz Biotechnology, Inc.), PERK (1:250; catalog no. sc-13073, lot no. RRID:AB_2230863; Santa Cruz Biotechnology, Inc.), IRE1α (1:500; catalog no. sc-20790, lot no. RRID:AB_2098712; Santa Cruz Biotechnology, Inc.), ATF6 (1:1,000; catalog no. ab37149, lot no. RRID:AB_725571; Abcam), Beclin1 (1:1,000; catalog no. sc-11427, lot no. RRID:AB_2064465; Santa Cruz Biotechnology, Inc.), ATG5 (1:1,000; catalog no. NB110-53818, lot no. RRID:AB_828587, Novus Biologicals), p62 (1:1,000; catalog no. 50–507-55, lot no. RRID: AB_2571590; Thermo Fisher Scientific), LC3 (1:2,000; catalog no. NB100-2220, lot no. RRID:AB_10003146; Novus Biologicals), and β-actin (1:5,000; catalog no. A1978, lot no. RRID:AB_476692; Sigma-Aldrich).

Techniques: Western Blot, Injection, Saline, Expressing, Staining, Plasmid Preparation, In Vitro, Control

Morphine-mediated autophagy in hippocampal neurons: Role of oxidative and ER stress. Representative Western blot (A) and quantification of autophagy mediators (Beclin1, ATG5, p62, and LC3-II; B) in the cell lysates of neurons pretreated with PBN or apocynin for 1 h followed by 24-h morphine treatment. (C) H 2 DCFDA assay of neurons pretreated with 3-MA (50 µM) for 30 min followed by morphine exposure. Representative Western blot (D) and quantification of Beclin1, ATG5, p62, and LC3-II (E) in the cell lysates of neurons pretreated with 4-PBA or salubrinal (100 µM) for 1 h followed by 24-h morphine exposure. Representative Western blot (F) and quantification of BIP, pPERK, IRE1α, and ATF6 (G) in the cell lysates of neurons pretreated with wortmannin or 3-MA for 1 h followed by 24-h morphine exposure. Quantification of Western blot results were all normalized to β-actin, except that pPERK was normalized to PERK. Each set of our results was quantified upon four independent experiments. All data are presented as mean ± SD. *, P < 0.05; **, P < 0.01 versus control/saline group; # , P < 0.05; ## , P < 0.01 versus morphine group using one-way ANOVA with post hoc test.

Journal: The Journal of Cell Biology

Article Title: Regulation of morphine-induced synaptic alterations: Role of oxidative stress, ER stress, and autophagy

doi: 10.1083/jcb.201605065

Figure Lengend Snippet: Morphine-mediated autophagy in hippocampal neurons: Role of oxidative and ER stress. Representative Western blot (A) and quantification of autophagy mediators (Beclin1, ATG5, p62, and LC3-II; B) in the cell lysates of neurons pretreated with PBN or apocynin for 1 h followed by 24-h morphine treatment. (C) H 2 DCFDA assay of neurons pretreated with 3-MA (50 µM) for 30 min followed by morphine exposure. Representative Western blot (D) and quantification of Beclin1, ATG5, p62, and LC3-II (E) in the cell lysates of neurons pretreated with 4-PBA or salubrinal (100 µM) for 1 h followed by 24-h morphine exposure. Representative Western blot (F) and quantification of BIP, pPERK, IRE1α, and ATF6 (G) in the cell lysates of neurons pretreated with wortmannin or 3-MA for 1 h followed by 24-h morphine exposure. Quantification of Western blot results were all normalized to β-actin, except that pPERK was normalized to PERK. Each set of our results was quantified upon four independent experiments. All data are presented as mean ± SD. *, P < 0.05; **, P < 0.01 versus control/saline group; # , P < 0.05; ## , P < 0.01 versus morphine group using one-way ANOVA with post hoc test.

Article Snippet: The Western blots were then probed with respective antibodies recognizing the vGlut1 (1:5,000; catalog no. AB5905, lot no. RRID:AB_2301751; EMD Millipore), PSD95 (1:1,000; catalog no. ab18258; lot no. RRID:AB_444362; Abcam), GAD65 (1:1,000; catalog no. NBP1-33284, lot no. RRID:AB_10004060; Novus Biologicals), gephyrin (1:250; catalog no. 610584, lot no. RRID:AB_397929; BD), BIP (1:1,000; catalog no. 610978, lot no. RRID:AB_398291; BD), pPERK (1:250; catalog no. sc-32577, lot no. RRID:AB_2293243; Santa Cruz Biotechnology, Inc.), PERK (1:250; catalog no. sc-13073, lot no. RRID:AB_2230863; Santa Cruz Biotechnology, Inc.), IRE1α (1:500; catalog no. sc-20790, lot no. RRID:AB_2098712; Santa Cruz Biotechnology, Inc.), ATF6 (1:1,000; catalog no. ab37149, lot no. RRID:AB_725571; Abcam), Beclin1 (1:1,000; catalog no. sc-11427, lot no. RRID:AB_2064465; Santa Cruz Biotechnology, Inc.), ATG5 (1:1,000; catalog no. NB110-53818, lot no. RRID:AB_828587, Novus Biologicals), p62 (1:1,000; catalog no. 50–507-55, lot no. RRID: AB_2571590; Thermo Fisher Scientific), LC3 (1:2,000; catalog no. NB100-2220, lot no. RRID:AB_10003146; Novus Biologicals), and β-actin (1:5,000; catalog no. A1978, lot no. RRID:AB_476692; Sigma-Aldrich).

Techniques: Western Blot, Control, Saline

Protective role of PDGF-BB in morphine-mediated synaptic alterations. Representative confocal images of GFP-expressing primary rat hippocampal neurons exposed to morphine followed by treatment with PDGF-BB (20 ng/ml) and stained with vGlut1 and GAD65 (A) and quantifications of spine density and excitatory and inhibitory synapses (B). Bar, 5 µm. Representative traces of whole-cell voltage-clamp recording showing mEPSC (C), mIPSC (E), mean mEPSC frequencies (D), and mean frequencies of mIPSCs (Hz; F) in primary rat neurons (DIV 19–21) treated with combinations of saline, morphine, and PDGF-BB as indicated. (G) ROS generation in neurons exposed to morphine and treated with PDGF-BB. Representative Western blot (H) and quantification of BIP, pPERK, IRE1α, and ATF6 (I) in the cell lysates of neurons exposed to morphine followed by treatment with PDGF-BB (20 ng/ml) for an additional 24 h. Representative Western blot (J) and quantification of Beclin1, ATG5, p62, and LC3-II (K) in the cell lysates of neurons exposed to morphine for 24 h followed by PDGF-BB (20 ng/ml) for additional 24 h. Quantification of Western blot results were all normalized to β-actin except that pPERK was normalized to PERK. Each set of our results was quantified upon four independent experiments. All data are presented as mean ± SD. *, P < 0.05; **, P < 0.01 versus control/saline group; # , P < 0.05; ## , P < 0.01 versus morphine group using one-way ANOVA with post hoc test.

Journal: The Journal of Cell Biology

Article Title: Regulation of morphine-induced synaptic alterations: Role of oxidative stress, ER stress, and autophagy

doi: 10.1083/jcb.201605065

Figure Lengend Snippet: Protective role of PDGF-BB in morphine-mediated synaptic alterations. Representative confocal images of GFP-expressing primary rat hippocampal neurons exposed to morphine followed by treatment with PDGF-BB (20 ng/ml) and stained with vGlut1 and GAD65 (A) and quantifications of spine density and excitatory and inhibitory synapses (B). Bar, 5 µm. Representative traces of whole-cell voltage-clamp recording showing mEPSC (C), mIPSC (E), mean mEPSC frequencies (D), and mean frequencies of mIPSCs (Hz; F) in primary rat neurons (DIV 19–21) treated with combinations of saline, morphine, and PDGF-BB as indicated. (G) ROS generation in neurons exposed to morphine and treated with PDGF-BB. Representative Western blot (H) and quantification of BIP, pPERK, IRE1α, and ATF6 (I) in the cell lysates of neurons exposed to morphine followed by treatment with PDGF-BB (20 ng/ml) for an additional 24 h. Representative Western blot (J) and quantification of Beclin1, ATG5, p62, and LC3-II (K) in the cell lysates of neurons exposed to morphine for 24 h followed by PDGF-BB (20 ng/ml) for additional 24 h. Quantification of Western blot results were all normalized to β-actin except that pPERK was normalized to PERK. Each set of our results was quantified upon four independent experiments. All data are presented as mean ± SD. *, P < 0.05; **, P < 0.01 versus control/saline group; # , P < 0.05; ## , P < 0.01 versus morphine group using one-way ANOVA with post hoc test.

Article Snippet: The Western blots were then probed with respective antibodies recognizing the vGlut1 (1:5,000; catalog no. AB5905, lot no. RRID:AB_2301751; EMD Millipore), PSD95 (1:1,000; catalog no. ab18258; lot no. RRID:AB_444362; Abcam), GAD65 (1:1,000; catalog no. NBP1-33284, lot no. RRID:AB_10004060; Novus Biologicals), gephyrin (1:250; catalog no. 610584, lot no. RRID:AB_397929; BD), BIP (1:1,000; catalog no. 610978, lot no. RRID:AB_398291; BD), pPERK (1:250; catalog no. sc-32577, lot no. RRID:AB_2293243; Santa Cruz Biotechnology, Inc.), PERK (1:250; catalog no. sc-13073, lot no. RRID:AB_2230863; Santa Cruz Biotechnology, Inc.), IRE1α (1:500; catalog no. sc-20790, lot no. RRID:AB_2098712; Santa Cruz Biotechnology, Inc.), ATF6 (1:1,000; catalog no. ab37149, lot no. RRID:AB_725571; Abcam), Beclin1 (1:1,000; catalog no. sc-11427, lot no. RRID:AB_2064465; Santa Cruz Biotechnology, Inc.), ATG5 (1:1,000; catalog no. NB110-53818, lot no. RRID:AB_828587, Novus Biologicals), p62 (1:1,000; catalog no. 50–507-55, lot no. RRID: AB_2571590; Thermo Fisher Scientific), LC3 (1:2,000; catalog no. NB100-2220, lot no. RRID:AB_10003146; Novus Biologicals), and β-actin (1:5,000; catalog no. A1978, lot no. RRID:AB_476692; Sigma-Aldrich).

Techniques: Expressing, Staining, Saline, Western Blot, Control

Exposure to a mixture of 7-oxysterols (2mix) causes lipid accumulation and autophagy dysfunction. THP-1 monocytes or THP-1 differentiated macrophages were treated with or without 2mix for 12 hours to 48 hours. In some experiments, cells were pretreated with the autophagy inhibitor 3MA or the autophagy inducer rapamycin for 1 hour and then exposed to 2mix or treated with the lysosomatropic agent CQ for 24 hours or CQ for 1 hour and 2mix for 24 hours. ( A ) Oil red O-stained THP-1 monocytes counterstained with hematoxylin (24 hours), bars = 100 µm. ( B and C ) Accumulation of autophagy vacuoles in THP-1 differentiated macrophages assayed by TEM and quantified by image analysis. ( B ) Photographs of low power view of control and 2mix-treated cells (two photographs on the left-hand side), bars = 2 µm or a higher magnified photograph (photograph on the top right-hand side) demonstrates autophagic vacuoles with double-membrane structure containing undigested cytoplasmic materials, bar = 1 µm. ( C ) Quantification of numbers of autophagic vacuoles, * P < 0.05, ** P < 0.01 versus control and 3MA + 2mix, respectively. ( D ) Atg5-immunostained THP-1 monocytes analyzed by flow cytometry. n = 6–8, * P < 0.05, ** P < 0.01 and *** P < 0.001. The inset shows a representative histogram of flow cytometry results (control: gray field histogram; 2mix: black-lined empty histogram). ( E ) LC3β-immunostained THP-1 monocytes analyzed by flow cytometry; n = 6–20, * P < 0.05. ( F ) Western blot analysis of LC3 I/II in the whole cell lysate of THP-1 monocytes. Values mentioned below the Western blot image are the ratios of LC3-II to LC3-I. ( G ) p62/ SQSTM1 -immunostained THP-1 monocytes analyzed by flow cytometry; n = 3–4. a P < 0.05 vs CQ and P < 0.01 vs 3MA + 2mix and CQ + 2mix. b P < 0.05 vs CQ and P < 0.01 vs CQ + 2mix and P < 0.001 vs 3MA + 2mix. * P < 0.05.

Journal: Journal of Cell Death

Article Title: Autophagy Induction Protects Against 7-Oxysterol-induced Cell Death via Lysosomal Pathway and Oxidative Stress

doi: 10.4137/JCD.S37841

Figure Lengend Snippet: Exposure to a mixture of 7-oxysterols (2mix) causes lipid accumulation and autophagy dysfunction. THP-1 monocytes or THP-1 differentiated macrophages were treated with or without 2mix for 12 hours to 48 hours. In some experiments, cells were pretreated with the autophagy inhibitor 3MA or the autophagy inducer rapamycin for 1 hour and then exposed to 2mix or treated with the lysosomatropic agent CQ for 24 hours or CQ for 1 hour and 2mix for 24 hours. ( A ) Oil red O-stained THP-1 monocytes counterstained with hematoxylin (24 hours), bars = 100 µm. ( B and C ) Accumulation of autophagy vacuoles in THP-1 differentiated macrophages assayed by TEM and quantified by image analysis. ( B ) Photographs of low power view of control and 2mix-treated cells (two photographs on the left-hand side), bars = 2 µm or a higher magnified photograph (photograph on the top right-hand side) demonstrates autophagic vacuoles with double-membrane structure containing undigested cytoplasmic materials, bar = 1 µm. ( C ) Quantification of numbers of autophagic vacuoles, * P < 0.05, ** P < 0.01 versus control and 3MA + 2mix, respectively. ( D ) Atg5-immunostained THP-1 monocytes analyzed by flow cytometry. n = 6–8, * P < 0.05, ** P < 0.01 and *** P < 0.001. The inset shows a representative histogram of flow cytometry results (control: gray field histogram; 2mix: black-lined empty histogram). ( E ) LC3β-immunostained THP-1 monocytes analyzed by flow cytometry; n = 6–20, * P < 0.05. ( F ) Western blot analysis of LC3 I/II in the whole cell lysate of THP-1 monocytes. Values mentioned below the Western blot image are the ratios of LC3-II to LC3-I. ( G ) p62/ SQSTM1 -immunostained THP-1 monocytes analyzed by flow cytometry; n = 3–4. a P < 0.05 vs CQ and P < 0.01 vs 3MA + 2mix and CQ + 2mix. b P < 0.05 vs CQ and P < 0.01 vs CQ + 2mix and P < 0.001 vs 3MA + 2mix. * P < 0.05.

Article Snippet: Different groups of cells were collected, fixed with 2% paraformaldehyde, permeabilized with 0.1% saponin in PBS, and incubated overnight with the following rabbit primary antibodies at 4°C: anti-LC3β antibody (Santa Cruz Biotechnology Inc.) or anti-Atg5 (Thermo Scientific) or anti-p62/ SQSTM1 (Medical and Biological Laboratories Co., Ltd).

Techniques: Staining, Flow Cytometry, Western Blot