sirna against atg7  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc sirna against atg7
    Effect of PE5 on autophagy regulatory proteins and autophagic cell death. ( A , B ) H460 cells were treated with PE5 and detected for LC3, p62, ATG5, and <t>ATG7</t> proteins by western blotting. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. ( C ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) and treated with PE5 for 24 h. Expression of LC3 was analyzed by immunofluorescence staining. ( D ) H460 cells were treated with PE5 in the presence of wortmannin (1 µM) or rapamycin (200 nM). Cell viability was analyzed by MTT assay. ( E , F ) Cells were transfected with <t>siATG7</t> and treated with 50 µM PE5 for 24 h. Expression levels of each ATG and LC3 were assessed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Cell viability was assessed using the MTT assay at 48 h. ( G ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) or Z-VAD-FMK (20 μM) (apoptosis inhibitor) and treated with PE5 for 24 h. Expression of PARP and cleaved PARP were analyzed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Data represent the mean ± SD (n = 3) and (* p < 0.05, ** p < 0.01, compared with the untreated control) ( # p < 0.05, compared with PE5-treated alone and significantly different from siATG-transfected cells).
    Sirna Against Atg7, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sirna against atg7/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sirna against atg7 - by Bioz Stars, 2023-03
    93/100 stars

    Images

    1) Product Images from "Targeting AKT/mTOR and Bcl-2 for Autophagic and Apoptosis Cell Death in Lung Cancer: Novel Activity of a Polyphenol Compound"

    Article Title: Targeting AKT/mTOR and Bcl-2 for Autophagic and Apoptosis Cell Death in Lung Cancer: Novel Activity of a Polyphenol Compound

    Journal: Antioxidants

    doi: 10.3390/antiox10040534

    Effect of PE5 on autophagy regulatory proteins and autophagic cell death. ( A , B ) H460 cells were treated with PE5 and detected for LC3, p62, ATG5, and ATG7 proteins by western blotting. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. ( C ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) and treated with PE5 for 24 h. Expression of LC3 was analyzed by immunofluorescence staining. ( D ) H460 cells were treated with PE5 in the presence of wortmannin (1 µM) or rapamycin (200 nM). Cell viability was analyzed by MTT assay. ( E , F ) Cells were transfected with siATG7 and treated with 50 µM PE5 for 24 h. Expression levels of each ATG and LC3 were assessed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Cell viability was assessed using the MTT assay at 48 h. ( G ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) or Z-VAD-FMK (20 μM) (apoptosis inhibitor) and treated with PE5 for 24 h. Expression of PARP and cleaved PARP were analyzed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Data represent the mean ± SD (n = 3) and (* p < 0.05, ** p < 0.01, compared with the untreated control) ( # p < 0.05, compared with PE5-treated alone and significantly different from siATG-transfected cells).
    Figure Legend Snippet: Effect of PE5 on autophagy regulatory proteins and autophagic cell death. ( A , B ) H460 cells were treated with PE5 and detected for LC3, p62, ATG5, and ATG7 proteins by western blotting. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. ( C ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) and treated with PE5 for 24 h. Expression of LC3 was analyzed by immunofluorescence staining. ( D ) H460 cells were treated with PE5 in the presence of wortmannin (1 µM) or rapamycin (200 nM). Cell viability was analyzed by MTT assay. ( E , F ) Cells were transfected with siATG7 and treated with 50 µM PE5 for 24 h. Expression levels of each ATG and LC3 were assessed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Cell viability was assessed using the MTT assay at 48 h. ( G ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) or Z-VAD-FMK (20 μM) (apoptosis inhibitor) and treated with PE5 for 24 h. Expression of PARP and cleaved PARP were analyzed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Data represent the mean ± SD (n = 3) and (* p < 0.05, ** p < 0.01, compared with the untreated control) ( # p < 0.05, compared with PE5-treated alone and significantly different from siATG-transfected cells).

    Techniques Used: Western Blot, Expressing, Immunofluorescence, Staining, MTT Assay, Transfection

    sirna against atg7  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc sirna against atg7
    Effect of PE5 on autophagy regulatory proteins and autophagic cell death. ( A , B ) H460 cells were treated with PE5 and detected for LC3, p62, ATG5, and <t>ATG7</t> proteins by western blotting. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. ( C ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) and treated with PE5 for 24 h. Expression of LC3 was analyzed by immunofluorescence staining. ( D ) H460 cells were treated with PE5 in the presence of wortmannin (1 µM) or rapamycin (200 nM). Cell viability was analyzed by MTT assay. ( E , F ) Cells were transfected with <t>siATG7</t> and treated with 50 µM PE5 for 24 h. Expression levels of each ATG and LC3 were assessed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Cell viability was assessed using the MTT assay at 48 h. ( G ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) or Z-VAD-FMK (20 μM) (apoptosis inhibitor) and treated with PE5 for 24 h. Expression of PARP and cleaved PARP were analyzed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Data represent the mean ± SD (n = 3) and (* p < 0.05, ** p < 0.01, compared with the untreated control) ( # p < 0.05, compared with PE5-treated alone and significantly different from siATG-transfected cells).
    Sirna Against Atg7, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sirna against atg7/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sirna against atg7 - by Bioz Stars, 2023-03
    93/100 stars

    Images

    1) Product Images from "Targeting AKT/mTOR and Bcl-2 for Autophagic and Apoptosis Cell Death in Lung Cancer: Novel Activity of a Polyphenol Compound"

    Article Title: Targeting AKT/mTOR and Bcl-2 for Autophagic and Apoptosis Cell Death in Lung Cancer: Novel Activity of a Polyphenol Compound

    Journal: Antioxidants

    doi: 10.3390/antiox10040534

    Effect of PE5 on autophagy regulatory proteins and autophagic cell death. ( A , B ) H460 cells were treated with PE5 and detected for LC3, p62, ATG5, and ATG7 proteins by western blotting. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. ( C ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) and treated with PE5 for 24 h. Expression of LC3 was analyzed by immunofluorescence staining. ( D ) H460 cells were treated with PE5 in the presence of wortmannin (1 µM) or rapamycin (200 nM). Cell viability was analyzed by MTT assay. ( E , F ) Cells were transfected with siATG7 and treated with 50 µM PE5 for 24 h. Expression levels of each ATG and LC3 were assessed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Cell viability was assessed using the MTT assay at 48 h. ( G ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) or Z-VAD-FMK (20 μM) (apoptosis inhibitor) and treated with PE5 for 24 h. Expression of PARP and cleaved PARP were analyzed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Data represent the mean ± SD (n = 3) and (* p < 0.05, ** p < 0.01, compared with the untreated control) ( # p < 0.05, compared with PE5-treated alone and significantly different from siATG-transfected cells).
    Figure Legend Snippet: Effect of PE5 on autophagy regulatory proteins and autophagic cell death. ( A , B ) H460 cells were treated with PE5 and detected for LC3, p62, ATG5, and ATG7 proteins by western blotting. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. ( C ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) and treated with PE5 for 24 h. Expression of LC3 was analyzed by immunofluorescence staining. ( D ) H460 cells were treated with PE5 in the presence of wortmannin (1 µM) or rapamycin (200 nM). Cell viability was analyzed by MTT assay. ( E , F ) Cells were transfected with siATG7 and treated with 50 µM PE5 for 24 h. Expression levels of each ATG and LC3 were assessed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Cell viability was assessed using the MTT assay at 48 h. ( G ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) or Z-VAD-FMK (20 μM) (apoptosis inhibitor) and treated with PE5 for 24 h. Expression of PARP and cleaved PARP were analyzed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Data represent the mean ± SD (n = 3) and (* p < 0.05, ** p < 0.01, compared with the untreated control) ( # p < 0.05, compared with PE5-treated alone and significantly different from siATG-transfected cells).

    Techniques Used: Western Blot, Expressing, Immunofluorescence, Staining, MTT Assay, Transfection

    sirna against atg7  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc sirna against atg7
    Altered energy homeostasis in mice lacking autophagy in SF1 neurons. (A) Body weight and (B) average food intake of <t>Atg7</t> loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n 4–6 per group) male mice. (C) Food intake of refed of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice after fasting (n = 5 for each group). (D) Body composition and (E) leptin sensitivity of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 5–6 for each group). (F) Serum leptin levels in fed and 12 h-fasted adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n = 3–5 per group) male mice. (G–I) Oxygen (O2) consumption and (J–L) carbon dioxide (CO2) production in (G, J) fed and (H, K) 12 h-fasted Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 5 for each group). Values are shown as mean ± SEM. ∗ P ≤ 0.05, and P ≤ 0.01 versus each group.
    Sirna Against Atg7, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sirna against atg7 - by Bioz Stars, 2023-03
    93/100 stars

    Images

    1) Product Images from "Defective autophagy in Sf1 neurons perturbs the metabolic response to fasting and causes mitochondrial dysfunction"

    Article Title: Defective autophagy in Sf1 neurons perturbs the metabolic response to fasting and causes mitochondrial dysfunction

    Journal: Molecular Metabolism

    doi: 10.1016/j.molmet.2021.101186

    Altered energy homeostasis in mice lacking autophagy in SF1 neurons. (A) Body weight and (B) average food intake of Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n 4–6 per group) male mice. (C) Food intake of refed of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice after fasting (n = 5 for each group). (D) Body composition and (E) leptin sensitivity of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 5–6 for each group). (F) Serum leptin levels in fed and 12 h-fasted adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n = 3–5 per group) male mice. (G–I) Oxygen (O2) consumption and (J–L) carbon dioxide (CO2) production in (G, J) fed and (H, K) 12 h-fasted Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 5 for each group). Values are shown as mean ± SEM. ∗ P ≤ 0.05, and P ≤ 0.01 versus each group.
    Figure Legend Snippet: Altered energy homeostasis in mice lacking autophagy in SF1 neurons. (A) Body weight and (B) average food intake of Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n 4–6 per group) male mice. (C) Food intake of refed of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice after fasting (n = 5 for each group). (D) Body composition and (E) leptin sensitivity of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 5–6 for each group). (F) Serum leptin levels in fed and 12 h-fasted adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n = 3–5 per group) male mice. (G–I) Oxygen (O2) consumption and (J–L) carbon dioxide (CO2) production in (G, J) fed and (H, K) 12 h-fasted Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 5 for each group). Values are shown as mean ± SEM. ∗ P ≤ 0.05, and P ≤ 0.01 versus each group.

    Techniques Used:

    Glucose metabolism in Sf1 -Cre; Atg7 loxP/loxP mice. (A) Serum glucose and (B) insulin levels and (C) VMH insulin receptor (InsR) mRNA expression in fed and 12h-fasted adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n = 4–7 per group) male mice. (D) Glucose and (E) insulin tolerance tests of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n = 4–6 per group) male mice. Values are shown as mean ± SEM. ∗ P ≤ 0.05, ∗∗ P ≤ 0.01, and ∗∗ P ≤ 0.001 versus each group.
    Figure Legend Snippet: Glucose metabolism in Sf1 -Cre; Atg7 loxP/loxP mice. (A) Serum glucose and (B) insulin levels and (C) VMH insulin receptor (InsR) mRNA expression in fed and 12h-fasted adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n = 4–7 per group) male mice. (D) Glucose and (E) insulin tolerance tests of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n = 4–6 per group) male mice. Values are shown as mean ± SEM. ∗ P ≤ 0.05, ∗∗ P ≤ 0.01, and ∗∗ P ≤ 0.001 versus each group.

    Techniques Used: Expressing

    Fasting-induced hypothalamic neuronal activation is impaired in Sf1 -Cre; Atg7 loxP/loxP mice. (A–C) Representative images and quantification of cFos-immunoreactive cells in the (A) ventromedial (VMH), (B) arcuate (ARH), and (C) dorsomedial (DMH) nuclei of the hypothalamus of fed and fasted adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 3–7 per group). (D) Vglut2 , (E) Pomc, and (F) Npy mRNA expression in the ARH of adult fed and fasted Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 3–4 per group). Values are shown as mean ± SEM. ∗ P ≤ 0.05 versus each group.
    Figure Legend Snippet: Fasting-induced hypothalamic neuronal activation is impaired in Sf1 -Cre; Atg7 loxP/loxP mice. (A–C) Representative images and quantification of cFos-immunoreactive cells in the (A) ventromedial (VMH), (B) arcuate (ARH), and (C) dorsomedial (DMH) nuclei of the hypothalamus of fed and fasted adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 3–7 per group). (D) Vglut2 , (E) Pomc, and (F) Npy mRNA expression in the ARH of adult fed and fasted Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 3–4 per group). Values are shown as mean ± SEM. ∗ P ≤ 0.05 versus each group.

    Techniques Used: Activation Assay, Expressing

    Loss of autophagy in SF1 neurons causes altered mitochondrial morphology and function. (A) Representative electron microscopy images of VMH neurons, percentage of autophagy-mitochondria contacts, and mitochondria aspect ratio in adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 4 per group). (B) Representative images and quantification of MitoTracker labeling in primary cultures of hypothalamic neurons 1 day after transfection with Atg7 siRNA or scrambled siRNA (n = 3–4 per group). (C) Mitochondria-related gene mRNA levels in the VMH of adult (10-week-old) fed or 12 h-fasted mice (n = 3–5 per group). (D) Measurement of mitochondrial respiration in the mediobasal hypothalamus of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 4–8 per group). Values are shown as mean ± SEM. ∗ P ≤ 0.05 versus each group.
    Figure Legend Snippet: Loss of autophagy in SF1 neurons causes altered mitochondrial morphology and function. (A) Representative electron microscopy images of VMH neurons, percentage of autophagy-mitochondria contacts, and mitochondria aspect ratio in adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 4 per group). (B) Representative images and quantification of MitoTracker labeling in primary cultures of hypothalamic neurons 1 day after transfection with Atg7 siRNA or scrambled siRNA (n = 3–4 per group). (C) Mitochondria-related gene mRNA levels in the VMH of adult (10-week-old) fed or 12 h-fasted mice (n = 3–5 per group). (D) Measurement of mitochondrial respiration in the mediobasal hypothalamus of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 4–8 per group). Values are shown as mean ± SEM. ∗ P ≤ 0.05 versus each group.

    Techniques Used: Electron Microscopy, Labeling, Transfection

    sirna against atg7  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc sirna against atg7
    Representative images and quantification of (A) ubiquitin- and (B) p62-immunoreactivity in the ventromedial nucleus of the hypothalamus (VMH) of adult <t>Atg7</t> loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 2-3 per group). ARH, arcuate nucleus of the hypothalamus; V3, third ventricle. Scale bar, 50 μm. Values are shown as mean ± SEM. * P ≤ 0.05 versus Atg7 loxP/loxP .
    Sirna Against Atg7, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sirna against atg7/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sirna against atg7 - by Bioz Stars, 2023-03
    93/100 stars

    Images

    1) Product Images from "Defective Autophagy in Sf1 Neurons Perturbs the Metabolic Response to Fasting and Causes Mitochondrial Dysfunction"

    Article Title: Defective Autophagy in Sf1 Neurons Perturbs the Metabolic Response to Fasting and Causes Mitochondrial Dysfunction

    Journal: bioRxiv

    doi: 10.1101/2020.11.02.348789

    Representative images and quantification of (A) ubiquitin- and (B) p62-immunoreactivity in the ventromedial nucleus of the hypothalamus (VMH) of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 2-3 per group). ARH, arcuate nucleus of the hypothalamus; V3, third ventricle. Scale bar, 50 μm. Values are shown as mean ± SEM. * P ≤ 0.05 versus Atg7 loxP/loxP .
    Figure Legend Snippet: Representative images and quantification of (A) ubiquitin- and (B) p62-immunoreactivity in the ventromedial nucleus of the hypothalamus (VMH) of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 2-3 per group). ARH, arcuate nucleus of the hypothalamus; V3, third ventricle. Scale bar, 50 μm. Values are shown as mean ± SEM. * P ≤ 0.05 versus Atg7 loxP/loxP .

    Techniques Used:

    (A) Body weight and (B) average food intake of Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n 4-6 per group) male mice. (C) Food intake of refed of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice after fasting (n = 5 for each group). (D) Body composition and (E) leptin sensitivity of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 5-6 for each group). (F) Serum leptin levels in fed and 12h-fasted adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n = 3-5 per group) male mice. (G-I) Oxygen (O2) consumption and (J-L) carbon dioxide (CO2) production in (G, J) fed and (H, K) 12h-fasted Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 5 for each group). Values are shown as mean ± SEM. * P ≤ 0.05, and P ≤ 0.01 versus each group.
    Figure Legend Snippet: (A) Body weight and (B) average food intake of Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n 4-6 per group) male mice. (C) Food intake of refed of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice after fasting (n = 5 for each group). (D) Body composition and (E) leptin sensitivity of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 5-6 for each group). (F) Serum leptin levels in fed and 12h-fasted adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n = 3-5 per group) male mice. (G-I) Oxygen (O2) consumption and (J-L) carbon dioxide (CO2) production in (G, J) fed and (H, K) 12h-fasted Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 5 for each group). Values are shown as mean ± SEM. * P ≤ 0.05, and P ≤ 0.01 versus each group.

    Techniques Used:

    (A-C) Respiratory quotient and (D-F) heat production in (A, D) fed and (B, E) 12h-fasted Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 4 for each group). Values are shown as mean ± SEM. * P ≤ 0.05 versus each group.
    Figure Legend Snippet: (A-C) Respiratory quotient and (D-F) heat production in (A, D) fed and (B, E) 12h-fasted Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 4 for each group). Values are shown as mean ± SEM. * P ≤ 0.05 versus each group.

    Techniques Used:

    (A) Serum glucose and (B) insulin levels, and (C) VMH insulin receptor (InsR) mRNA expression in fed and 12h-fasted adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n = 4-7 per group) male mice. (D) Glucose and (E) insulin tolerance tests and area under the curves of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n = 4-6 per group) male mice. Values are shown as mean ± SEM. * P ≤ 0.05, ** P ≤ 0.01, and ** P ≤ 0.001 versus each group.
    Figure Legend Snippet: (A) Serum glucose and (B) insulin levels, and (C) VMH insulin receptor (InsR) mRNA expression in fed and 12h-fasted adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n = 4-7 per group) male mice. (D) Glucose and (E) insulin tolerance tests and area under the curves of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP (n = 4-6 per group) male mice. Values are shown as mean ± SEM. * P ≤ 0.05, ** P ≤ 0.01, and ** P ≤ 0.001 versus each group.

    Techniques Used: Expressing

    (A-C) Representative images and quantification of cFos-immunoreactive cells in the (A) ventromedial (VMH), (B) arcuate (ARH), and (C) dorsomedial (DMH) nuclei of the hypothalamus of fed and fasted adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 3-7 per group). (D) Vglut2 , (E) Pomc, and (F) Npy mRNA expression in the ARH of adult fed and fasted Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 3-4 per group). Values are shown as mean ± SEM. * P ≤ 0.05 versus each group.
    Figure Legend Snippet: (A-C) Representative images and quantification of cFos-immunoreactive cells in the (A) ventromedial (VMH), (B) arcuate (ARH), and (C) dorsomedial (DMH) nuclei of the hypothalamus of fed and fasted adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 3-7 per group). (D) Vglut2 , (E) Pomc, and (F) Npy mRNA expression in the ARH of adult fed and fasted Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 3-4 per group). Values are shown as mean ± SEM. * P ≤ 0.05 versus each group.

    Techniques Used: Expressing

    (A) Representative electron microscopy images of VMH neurons, percentage of autophagy-mitochondria contacts, and mitochondria aspect ratio in adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n= 4 per group). (B) Representative images and quantification of MitoTracker labeling in primary cultures of hypothalamic neurons 1 day after transfection with Atg7 siRNA or scrambled siRNA (n = 3-4 per group). (C) Mitochondria-related gene mRNA levels in the VMH of adult (10-week-old) fed or 12h-fasted mice (n = 3-5 per group). (D) Measurement of mitochondrial respiration in the mediobasal hypothalamus of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 4-8 per group). Values are shown as mean ± SEM. * P ≤ 0.05 versus each group.
    Figure Legend Snippet: (A) Representative electron microscopy images of VMH neurons, percentage of autophagy-mitochondria contacts, and mitochondria aspect ratio in adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n= 4 per group). (B) Representative images and quantification of MitoTracker labeling in primary cultures of hypothalamic neurons 1 day after transfection with Atg7 siRNA or scrambled siRNA (n = 3-4 per group). (C) Mitochondria-related gene mRNA levels in the VMH of adult (10-week-old) fed or 12h-fasted mice (n = 3-5 per group). (D) Measurement of mitochondrial respiration in the mediobasal hypothalamus of adult Atg7 loxP/loxP and Sf1 -Cre; Atg7 loxP/loxP male mice (n = 4-8 per group). Values are shown as mean ± SEM. * P ≤ 0.05 versus each group.

    Techniques Used: Electron Microscopy, Labeling, Transfection

    sirna against atg7  (Cell Signaling Technology Inc)


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    sirna against atg7  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc sirna against atg7
    Effect of PE5 on autophagy regulatory proteins and autophagic cell death. ( A , B ) H460 cells were treated with PE5 and detected for LC3, p62, ATG5, and <t>ATG7</t> proteins by western blotting. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. ( C ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) and treated with PE5 for 24 h. Expression of LC3 was analyzed by immunofluorescence staining. ( D ) H460 cells were treated with PE5 in the presence of wortmannin (1 µM) or rapamycin (200 nM). Cell viability was analyzed by MTT assay. ( E , F ) Cells were transfected with <t>siATG7</t> and treated with 50 µM PE5 for 24 h. Expression levels of each ATG and LC3 were assessed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Cell viability was assessed using the MTT assay at 48 h. ( G ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) or Z-VAD-FMK (20 μM) (apoptosis inhibitor) and treated with PE5 for 24 h. Expression of PARP and cleaved PARP were analyzed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Data represent the mean ± SD (n = 3) and (* p < 0.05, ** p < 0.01, compared with the untreated control) ( # p < 0.05, compared with PE5-treated alone and significantly different from siATG-transfected cells).
    Sirna Against Atg7, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Effect of PE5 on autophagy regulatory proteins and autophagic cell death. ( A , B ) H460 cells were treated with PE5 and detected for LC3, p62, ATG5, and ATG7 proteins by western blotting. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. ( C ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) and treated with PE5 for 24 h. Expression of LC3 was analyzed by immunofluorescence staining. ( D ) H460 cells were treated with PE5 in the presence of wortmannin (1 µM) or rapamycin (200 nM). Cell viability was analyzed by MTT assay. ( E , F ) Cells were transfected with siATG7 and treated with 50 µM PE5 for 24 h. Expression levels of each ATG and LC3 were assessed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Cell viability was assessed using the MTT assay at 48 h. ( G ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) or Z-VAD-FMK (20 μM) (apoptosis inhibitor) and treated with PE5 for 24 h. Expression of PARP and cleaved PARP were analyzed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Data represent the mean ± SD (n = 3) and (* p < 0.05, ** p < 0.01, compared with the untreated control) ( # p < 0.05, compared with PE5-treated alone and significantly different from siATG-transfected cells).

    Journal: Antioxidants

    Article Title: Targeting AKT/mTOR and Bcl-2 for Autophagic and Apoptosis Cell Death in Lung Cancer: Novel Activity of a Polyphenol Compound

    doi: 10.3390/antiox10040534

    Figure Lengend Snippet: Effect of PE5 on autophagy regulatory proteins and autophagic cell death. ( A , B ) H460 cells were treated with PE5 and detected for LC3, p62, ATG5, and ATG7 proteins by western blotting. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. ( C ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) and treated with PE5 for 24 h. Expression of LC3 was analyzed by immunofluorescence staining. ( D ) H460 cells were treated with PE5 in the presence of wortmannin (1 µM) or rapamycin (200 nM). Cell viability was analyzed by MTT assay. ( E , F ) Cells were transfected with siATG7 and treated with 50 µM PE5 for 24 h. Expression levels of each ATG and LC3 were assessed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Cell viability was assessed using the MTT assay at 48 h. ( G ) H460 cells were pre-treated with wortmannin (1 µM) (an autophagic inhibitor) or Z-VAD-FMK (20 μM) (apoptosis inhibitor) and treated with PE5 for 24 h. Expression of PARP and cleaved PARP were analyzed by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. Data represent the mean ± SD (n = 3) and (* p < 0.05, ** p < 0.01, compared with the untreated control) ( # p < 0.05, compared with PE5-treated alone and significantly different from siATG-transfected cells).

    Article Snippet: The siRNA including siRNA control and siRNA against ATG7 were obtained from Cell Signaling Technology, Inc. (Danvers, MA, USA).

    Techniques: Western Blot, Expressing, Immunofluorescence, Staining, MTT Assay, Transfection