rabbit polyclonal antibody pampkα Search Results


rabbit anti pampk  (Cell Signaling Technology Inc)
96
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Rabbit Anti Pampk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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antibody against pampk  (Cell Signaling Technology Inc)
99
Cell Signaling Technology Inc antibody against pampk
AMPK phosphorylation and content in hypothalamus of rats exposed to fructose consumption during the light or the dark phases. Rats assigned to the groups control (CTL), Light Phase Fructose (LPF) and Dark Phase Fructose (DPF) had their hypothalamus removed at the end of the eighth week of treatment. A first set of samples was used for ( A ) Western blot detection of phosphorylated Adenosine Monophosphate-activated protein kinase (AMPK) and ( B ) total AMPK. Target proteins were normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH). A second set of samples was processed for immunofluorescent staining. Sections were stained using <t>an</t> <t>anti-pAMPK</t> antibody followed by secondary antibody conjugated to Alexafluor 546 (red). Nuclear structures are visualized by 4′,6-diamidino-2-phenylindole (DAPI) probing (Blue). ( C ) Large magnification (400×) images are shown from the arcuate nucleus (ARC), lateral hypothalamus (LH), ventro medial hypothalamus (VMH) and paraventricular nucleus (PVN). The results are presented as the means ± standard error of the mean. ** p < 0.05 vs. CTL.
Antibody Against Pampk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit polyclonal anti pampkα t172  (Cell Signaling Technology Inc)
97
Cell Signaling Technology Inc rabbit polyclonal anti pampkα t172
( A ) WT or FKBP51 KO cells were starved in HBSS medium for 4 hours to induce autophagy, followed by quantification of <t>pAMPKα</t> <t>(T172),</t> ( B ) p62, and ( C ) pp70S6K (T389). Representative blots are shown in ( D ). FKBP51 overexpression (FKBP51 OE) in N2a cells (see fig. S3D for validation) enhanced autophagy signaling. Quantification of ( E ) pAMPKα (T172), ( F ) pp70S6K (T389), ( G ) p62, and ( H ) representative blots. ( I ) Quantification of autophagic flux in FKBP51 KO and FKBP51 OE cells in response to starvation. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. ( J ) Representative blots of autophagic flux measurements. ( K ) Representative pictures of TFEB nuclear localization/translocation. DAPI, 4′,6-diamidino-2-phenylindole. Scale bar, 10 μm. ( L ) Quantification of TFEB reporter assay. BL, baseline. All data (A to J) are shown as relative fold change compared to control condition; ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001; ## P < 0.01, ### P < 0.001; $$ P < 0.01. Two-way ANOVA was performed in (A) to (C) and followed by a Tukey’s multiple comparisons test. One-way ANOVA was performed for (I) and (L), followed by a Dunnett’s multiple comparison test. The unpaired Student’s t test was performed for (E) to (G). *, significant genotype effect; $, significant starvation effect; #, significant treatment effect.
Rabbit Polyclonal Anti Pampkα T172, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti phosphorylated ampk p ampk  (Cell Signaling Technology Inc)
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Cell Signaling Technology Inc rabbit anti phosphorylated ampk p ampk
( A ) WT or FKBP51 KO cells were starved in HBSS medium for 4 hours to induce autophagy, followed by quantification of <t>pAMPKα</t> <t>(T172),</t> ( B ) p62, and ( C ) pp70S6K (T389). Representative blots are shown in ( D ). FKBP51 overexpression (FKBP51 OE) in N2a cells (see fig. S3D for validation) enhanced autophagy signaling. Quantification of ( E ) pAMPKα (T172), ( F ) pp70S6K (T389), ( G ) p62, and ( H ) representative blots. ( I ) Quantification of autophagic flux in FKBP51 KO and FKBP51 OE cells in response to starvation. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. ( J ) Representative blots of autophagic flux measurements. ( K ) Representative pictures of TFEB nuclear localization/translocation. DAPI, 4′,6-diamidino-2-phenylindole. Scale bar, 10 μm. ( L ) Quantification of TFEB reporter assay. BL, baseline. All data (A to J) are shown as relative fold change compared to control condition; ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001; ## P < 0.01, ### P < 0.001; $$ P < 0.01. Two-way ANOVA was performed in (A) to (C) and followed by a Tukey’s multiple comparisons test. One-way ANOVA was performed for (I) and (L), followed by a Dunnett’s multiple comparison test. The unpaired Student’s t test was performed for (E) to (G). *, significant genotype effect; $, significant starvation effect; #, significant treatment effect.
Rabbit Anti Phosphorylated Ampk P Ampk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti-human phospho-ampk antibody  (Santa Cruz Biotechnology)
90
Santa Cruz Biotechnology rabbit anti-human phospho-ampk antibody
( A ) WT or FKBP51 KO cells were starved in HBSS medium for 4 hours to induce autophagy, followed by quantification of <t>pAMPKα</t> <t>(T172),</t> ( B ) p62, and ( C ) pp70S6K (T389). Representative blots are shown in ( D ). FKBP51 overexpression (FKBP51 OE) in N2a cells (see fig. S3D for validation) enhanced autophagy signaling. Quantification of ( E ) pAMPKα (T172), ( F ) pp70S6K (T389), ( G ) p62, and ( H ) representative blots. ( I ) Quantification of autophagic flux in FKBP51 KO and FKBP51 OE cells in response to starvation. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. ( J ) Representative blots of autophagic flux measurements. ( K ) Representative pictures of TFEB nuclear localization/translocation. DAPI, 4′,6-diamidino-2-phenylindole. Scale bar, 10 μm. ( L ) Quantification of TFEB reporter assay. BL, baseline. All data (A to J) are shown as relative fold change compared to control condition; ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001; ## P < 0.01, ### P < 0.001; $$ P < 0.01. Two-way ANOVA was performed in (A) to (C) and followed by a Tukey’s multiple comparisons test. One-way ANOVA was performed for (I) and (L), followed by a Dunnett’s multiple comparison test. The unpaired Student’s t test was performed for (E) to (G). *, significant genotype effect; $, significant starvation effect; #, significant treatment effect.
Rabbit Anti Human Phospho Ampk Antibody, supplied by Santa Cruz Biotechnology, 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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phosphorylated ampk (pampk) primary antibody rabbit polyclonal  (Cell Signaling Technology Inc)
90
Cell Signaling Technology Inc phosphorylated ampk (pampk) primary antibody rabbit polyclonal
( A ) Representative Western blots of <t>phosphorylated</t> AMP-activated protein kinase <t>(pAMPK)</t> and total <t>AMPK</t> performed in TA muscles from sedentary mice and exercised mice treated with vehicle, BCAAs , mix 1 , mix 2 , or mix 3 . ( B ) Calculation of the pAMPK/AMPK ratio for each experimental group. Values are expressed as mean ± SEM from the number of mice indicated in brackets. No statistically significant differences between vehicle and treated groups were found by one-way ANOVA followed by Bonferroni post hoc correction. Individual comparisons of the means vs. sedentary mice showed statistically significant differences found by unpaired Student’s t -test, which are indicated above the bars.
Phosphorylated Ampk (Pampk) Primary Antibody Rabbit Polyclonal, supplied by Cell Signaling Technology Inc, 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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rabbit monoclonal pampk thr172  (Cell Signaling Technology Inc)
95
Cell Signaling Technology Inc rabbit monoclonal pampk thr172
( A ) Representative Western blots of <t>phosphorylated</t> AMP-activated protein kinase <t>(pAMPK)</t> and total <t>AMPK</t> performed in TA muscles from sedentary mice and exercised mice treated with vehicle, BCAAs , mix 1 , mix 2 , or mix 3 . ( B ) Calculation of the pAMPK/AMPK ratio for each experimental group. Values are expressed as mean ± SEM from the number of mice indicated in brackets. No statistically significant differences between vehicle and treated groups were found by one-way ANOVA followed by Bonferroni post hoc correction. Individual comparisons of the means vs. sedentary mice showed statistically significant differences found by unpaired Student’s t -test, which are indicated above the bars.
Rabbit Monoclonal Pampk Thr172, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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antibody rabbit anti-pampk  (Cell Signaling Technology Inc)
90
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( A ) Representative Western blots of <t>phosphorylated</t> AMP-activated protein kinase <t>(pAMPK)</t> and total <t>AMPK</t> performed in TA muscles from sedentary mice and exercised mice treated with vehicle, BCAAs , mix 1 , mix 2 , or mix 3 . ( B ) Calculation of the pAMPK/AMPK ratio for each experimental group. Values are expressed as mean ± SEM from the number of mice indicated in brackets. No statistically significant differences between vehicle and treated groups were found by one-way ANOVA followed by Bonferroni post hoc correction. Individual comparisons of the means vs. sedentary mice showed statistically significant differences found by unpaired Student’s t -test, which are indicated above the bars.
Antibody Rabbit Anti Pampk, supplied by Cell Signaling Technology Inc, 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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rabbit anti-pampk 44-1150g  (Thermo Fisher)
90
Thermo Fisher rabbit anti-pampk 44-1150g
( A ) Representative Western blots of <t>phosphorylated</t> AMP-activated protein kinase <t>(pAMPK)</t> and total <t>AMPK</t> performed in TA muscles from sedentary mice and exercised mice treated with vehicle, BCAAs , mix 1 , mix 2 , or mix 3 . ( B ) Calculation of the pAMPK/AMPK ratio for each experimental group. Values are expressed as mean ± SEM from the number of mice indicated in brackets. No statistically significant differences between vehicle and treated groups were found by one-way ANOVA followed by Bonferroni post hoc correction. Individual comparisons of the means vs. sedentary mice showed statistically significant differences found by unpaired Student’s t -test, which are indicated above the bars.
Rabbit Anti Pampk 44 1150g, 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
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rabbit monoclonal phosphoampk  (Cell Signaling Technology Inc)
98
Cell Signaling Technology Inc rabbit monoclonal phosphoampk
( A ) Representative Western blots of <t>phosphorylated</t> AMP-activated protein kinase <t>(pAMPK)</t> and total <t>AMPK</t> performed in TA muscles from sedentary mice and exercised mice treated with vehicle, BCAAs , mix 1 , mix 2 , or mix 3 . ( B ) Calculation of the pAMPK/AMPK ratio for each experimental group. Values are expressed as mean ± SEM from the number of mice indicated in brackets. No statistically significant differences between vehicle and treated groups were found by one-way ANOVA followed by Bonferroni post hoc correction. Individual comparisons of the means vs. sedentary mice showed statistically significant differences found by unpaired Student’s t -test, which are indicated above the bars.
Rabbit Monoclonal Phosphoampk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti pampka  (Cell Signaling Technology Inc)
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Cell Signaling Technology Inc anti pampka
KEY RESOURCES TABLE
Anti Pampka, supplied by Cell Signaling Technology Inc, 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/anti pampka/product/Cell Signaling Technology Inc
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anti pampkα t172  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc anti pampkα t172
KEY RESOURCES TABLE
Anti Pampkα T172, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


AMPK phosphorylation and content in hypothalamus of rats exposed to fructose consumption during the light or the dark phases. Rats assigned to the groups control (CTL), Light Phase Fructose (LPF) and Dark Phase Fructose (DPF) had their hypothalamus removed at the end of the eighth week of treatment. A first set of samples was used for ( A ) Western blot detection of phosphorylated Adenosine Monophosphate-activated protein kinase (AMPK) and ( B ) total AMPK. Target proteins were normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH). A second set of samples was processed for immunofluorescent staining. Sections were stained using an anti-pAMPK antibody followed by secondary antibody conjugated to Alexafluor 546 (red). Nuclear structures are visualized by 4′,6-diamidino-2-phenylindole (DAPI) probing (Blue). ( C ) Large magnification (400×) images are shown from the arcuate nucleus (ARC), lateral hypothalamus (LH), ventro medial hypothalamus (VMH) and paraventricular nucleus (PVN). The results are presented as the means ± standard error of the mean. ** p < 0.05 vs. CTL.

Journal: Nutrients

Article Title: Metabolic Impact of Light Phase-Restricted Fructose Consumption Is Linked to Changes in Hypothalamic AMPK Phosphorylation and Melatonin Production in Rats

doi: 10.3390/nu9040332

Figure Lengend Snippet: AMPK phosphorylation and content in hypothalamus of rats exposed to fructose consumption during the light or the dark phases. Rats assigned to the groups control (CTL), Light Phase Fructose (LPF) and Dark Phase Fructose (DPF) had their hypothalamus removed at the end of the eighth week of treatment. A first set of samples was used for ( A ) Western blot detection of phosphorylated Adenosine Monophosphate-activated protein kinase (AMPK) and ( B ) total AMPK. Target proteins were normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH). A second set of samples was processed for immunofluorescent staining. Sections were stained using an anti-pAMPK antibody followed by secondary antibody conjugated to Alexafluor 546 (red). Nuclear structures are visualized by 4′,6-diamidino-2-phenylindole (DAPI) probing (Blue). ( C ) Large magnification (400×) images are shown from the arcuate nucleus (ARC), lateral hypothalamus (LH), ventro medial hypothalamus (VMH) and paraventricular nucleus (PVN). The results are presented as the means ± standard error of the mean. ** p < 0.05 vs. CTL.

Article Snippet: Sections were incubated with primary antibody against pAMPK (Thr 172) (Cat. 2535, Cell Signaling Technology, Danvers, MA, USA) overnight at −4 °C and with secondary antibody conjugated to Alexafluor 546 for 2 h (Cat. A10040, Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Western Blot, Staining

( A ) WT or FKBP51 KO cells were starved in HBSS medium for 4 hours to induce autophagy, followed by quantification of pAMPKα (T172), ( B ) p62, and ( C ) pp70S6K (T389). Representative blots are shown in ( D ). FKBP51 overexpression (FKBP51 OE) in N2a cells (see fig. S3D for validation) enhanced autophagy signaling. Quantification of ( E ) pAMPKα (T172), ( F ) pp70S6K (T389), ( G ) p62, and ( H ) representative blots. ( I ) Quantification of autophagic flux in FKBP51 KO and FKBP51 OE cells in response to starvation. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. ( J ) Representative blots of autophagic flux measurements. ( K ) Representative pictures of TFEB nuclear localization/translocation. DAPI, 4′,6-diamidino-2-phenylindole. Scale bar, 10 μm. ( L ) Quantification of TFEB reporter assay. BL, baseline. All data (A to J) are shown as relative fold change compared to control condition; ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001; ## P < 0.01, ### P < 0.001; $$ P < 0.01. Two-way ANOVA was performed in (A) to (C) and followed by a Tukey’s multiple comparisons test. One-way ANOVA was performed for (I) and (L), followed by a Dunnett’s multiple comparison test. The unpaired Student’s t test was performed for (E) to (G). *, significant genotype effect; $, significant starvation effect; #, significant treatment effect.

Journal: Science Advances

Article Title: Mediobasal hypothalamic FKBP51 acts as a molecular switch linking autophagy to whole-body metabolism

doi: 10.1126/sciadv.abi4797

Figure Lengend Snippet: ( A ) WT or FKBP51 KO cells were starved in HBSS medium for 4 hours to induce autophagy, followed by quantification of pAMPKα (T172), ( B ) p62, and ( C ) pp70S6K (T389). Representative blots are shown in ( D ). FKBP51 overexpression (FKBP51 OE) in N2a cells (see fig. S3D for validation) enhanced autophagy signaling. Quantification of ( E ) pAMPKα (T172), ( F ) pp70S6K (T389), ( G ) p62, and ( H ) representative blots. ( I ) Quantification of autophagic flux in FKBP51 KO and FKBP51 OE cells in response to starvation. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. ( J ) Representative blots of autophagic flux measurements. ( K ) Representative pictures of TFEB nuclear localization/translocation. DAPI, 4′,6-diamidino-2-phenylindole. Scale bar, 10 μm. ( L ) Quantification of TFEB reporter assay. BL, baseline. All data (A to J) are shown as relative fold change compared to control condition; ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001; ## P < 0.01, ### P < 0.001; $$ P < 0.01. Two-way ANOVA was performed in (A) to (C) and followed by a Tukey’s multiple comparisons test. One-way ANOVA was performed for (I) and (L), followed by a Dunnett’s multiple comparison test. The unpaired Student’s t test was performed for (E) to (G). *, significant genotype effect; $, significant starvation effect; #, significant treatment effect.

Article Snippet: The following antibodies were used: goat polyclonal anti-actin (I-19) (sc-1616, Santa Cruz Biotechnology), rabbit polyclonal anti-FKBP51 (A301-430A, Bethyl Laboratories), rabbit monoclonal anti-FKBP5 (D5G2, #12210, Cell Signaling Technology), rabbit monoclonal anti-LKB1 (D60C5, #3047, Cell Signaling Technology), rabbit polyclonal anti-pAMPKα T172 (#2531, Cell Signaling Technology), rabbit polyclonal anti-pAMPKα (#2532, Cell Signaling Technology), rabbit polyclonal anti-SKP2 (L70, #4313, Cell Signaling Technology), rabbit anti-pSKP2 S72 (was a gift from Cell Signaling Technology), rabbit polyclonal anti-AKT (#9272, Cell Signaling Technology), rabbit monoclonal anti-pAKT S473 (D9E, #4060, Cell Signaling Technology), rabbit polyclonal anti-p62 (#5114, Cell Signaling Technology), rabbit monoclonal anti-LC3B (D11, #3868, Cell Signaling Technology), rabbit polyclonal anti-pULK1 S757 (#6888, Cell Signaling Technology), rabbit monoclonal anti-pULK1 S555 (D1H4, #5869, Cell Signaling Technology), rabbit monoclonal anti-ULK1 (D8H5, #8054, Cell Signaling Technology), anti-pBECN1 S93/S96 (in mouse S91/S94) (#12476, Cell Signaling Technology), rabbit polyclonal anti-pBECN1 S15 (#84966, Cell Signaling Technology), rabbit polyclonal anti-BECN1 (#3738, Cell Signaling Technology), rabbit polyclonal anti-TSC2 (#3612, Cell Signaling Technology), rabbit polyclonal anti-pTSC2 S1387 (#5584, Cell Signaling Technology), rabbit monoclonal anti-pATG16L1 S278 (EPR19016, ab195242, Abcam), rabbit polyclonal anti-WIPI4 (WDR45) (19194-1-AP, Proteintech), mouse monoclonal anti-WIPI4 (G12, sc-398272, Santa Cruz Biotechnology), rabbit polyclonal anti-WIPI3 (WDR45L) (SAB2102704, Sigma-Aldrich), mouse monoclonal anti-WIPI3 (B-7, sc-514194, Santa Cruz Biotechnology), rabbit polyclonal anti-WIPI2 (#8567, Cell Signaling Technology), rabbit polyclonal anti-WIPI1 (HPA007493, Sigma-Aldrich), rabbit polyclonal anti-AMPKα1 (#2795, Cell Signaling Technology), rabbit polyclonal anti-AMPKγ2 (#2536, Cell Signaling Technology), rabbit polyclonal anti-AMPKα2 (#2757, Cell Signaling Technology), rabbit monoclonal anti-AMPKβ1 (71C10, #4178, Cell Signaling Technology), rabbit polyclonal anti-AMPKγ1 (#4187, Cell Signaling Technology), rabbit polyclonal anti-AMPKβ2 (#4188, Cell Signaling Technology), rabbit polyclonal anti-AMPKγ3 (#2550, Cell Signaling Technology), rabbit monoclonal anti-TSC1 (D43E2, #6935, Cell Signaling Technology), rabbit polyclonal anti-Flag (600-401-383, Rockland Inc.), rabbit polyclonal anti-hypusine (ABS1046, Merck Millipore), rabbit monoclonal anti-eIF5A (D8L8Q, #20765, Cell Signaling Technology), and rabbit polyclonal anti-TFEB (ab245350, Abcam).

Techniques: Over Expression, Biomarker Discovery, Translocation Assay, Reporter Assay, Control, Comparison

FKBP51 deletion is depicted in green, and FKBP51 overexpression is depicted in blue. ( A ) Representative blots of autophagy and mTOR markers in FKBP51 MBH-KO mice. ( B ) Quantification of FKBP51 deletion. ( C ) FKBP51 deletion reduced LKB1 and AMPK binding to WIPI4 as well as ( D ) AMPK phosphorylation at T172. ( E ) TSC2-WIPI3 binding was decreased in FKBP51 MBH-KO animals. ( F ) Quantification of mTOR substrate pp70S6K (T389). ( G ) LC3B-II and ( H ) p62 levels in the MBH. ( I ) Representative blots of autophagy and mTOR marker in FKBP51 MBH-OE mice. ( J ) Quantification of viral FKBP51 overexpression. ( K ) FKBP51 overexpression reduced LKB1 and AMPK binding to WIPI4. ( L ) Quantification of AMPK phosphorylation at T172. ( M ) TSC2-WIPI3 binding was decreased. ( N ) Quantification of pp70S6K phosphorylation at T389. ( O ) To assess autophagic flux FKBP51MBH-OE, animals were treated with chloroquine (50 mg/kg), and LC3B-II levels were analyzed 4 hours after treatment. ( P ) FKBP51 overexpression blocked autophagic flux and resulted in an accumulation of p62. ( Q and R ) Quantification of FKBP51, p62, and BECN1, while titrating AAV-HA-FKBP51 virus into mouse neuroblastoma cells. ( S ) MBH FKBP51 regulates autophagy and mTOR signaling in a dose-dependent manner. All data are shown as ±SEM. Data are shown as the relative protein expression compared to control; for (A) to (N), an unpaired Student’s t test was performed. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Journal: Science Advances

Article Title: Mediobasal hypothalamic FKBP51 acts as a molecular switch linking autophagy to whole-body metabolism

doi: 10.1126/sciadv.abi4797

Figure Lengend Snippet: FKBP51 deletion is depicted in green, and FKBP51 overexpression is depicted in blue. ( A ) Representative blots of autophagy and mTOR markers in FKBP51 MBH-KO mice. ( B ) Quantification of FKBP51 deletion. ( C ) FKBP51 deletion reduced LKB1 and AMPK binding to WIPI4 as well as ( D ) AMPK phosphorylation at T172. ( E ) TSC2-WIPI3 binding was decreased in FKBP51 MBH-KO animals. ( F ) Quantification of mTOR substrate pp70S6K (T389). ( G ) LC3B-II and ( H ) p62 levels in the MBH. ( I ) Representative blots of autophagy and mTOR marker in FKBP51 MBH-OE mice. ( J ) Quantification of viral FKBP51 overexpression. ( K ) FKBP51 overexpression reduced LKB1 and AMPK binding to WIPI4. ( L ) Quantification of AMPK phosphorylation at T172. ( M ) TSC2-WIPI3 binding was decreased. ( N ) Quantification of pp70S6K phosphorylation at T389. ( O ) To assess autophagic flux FKBP51MBH-OE, animals were treated with chloroquine (50 mg/kg), and LC3B-II levels were analyzed 4 hours after treatment. ( P ) FKBP51 overexpression blocked autophagic flux and resulted in an accumulation of p62. ( Q and R ) Quantification of FKBP51, p62, and BECN1, while titrating AAV-HA-FKBP51 virus into mouse neuroblastoma cells. ( S ) MBH FKBP51 regulates autophagy and mTOR signaling in a dose-dependent manner. All data are shown as ±SEM. Data are shown as the relative protein expression compared to control; for (A) to (N), an unpaired Student’s t test was performed. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: The following antibodies were used: goat polyclonal anti-actin (I-19) (sc-1616, Santa Cruz Biotechnology), rabbit polyclonal anti-FKBP51 (A301-430A, Bethyl Laboratories), rabbit monoclonal anti-FKBP5 (D5G2, #12210, Cell Signaling Technology), rabbit monoclonal anti-LKB1 (D60C5, #3047, Cell Signaling Technology), rabbit polyclonal anti-pAMPKα T172 (#2531, Cell Signaling Technology), rabbit polyclonal anti-pAMPKα (#2532, Cell Signaling Technology), rabbit polyclonal anti-SKP2 (L70, #4313, Cell Signaling Technology), rabbit anti-pSKP2 S72 (was a gift from Cell Signaling Technology), rabbit polyclonal anti-AKT (#9272, Cell Signaling Technology), rabbit monoclonal anti-pAKT S473 (D9E, #4060, Cell Signaling Technology), rabbit polyclonal anti-p62 (#5114, Cell Signaling Technology), rabbit monoclonal anti-LC3B (D11, #3868, Cell Signaling Technology), rabbit polyclonal anti-pULK1 S757 (#6888, Cell Signaling Technology), rabbit monoclonal anti-pULK1 S555 (D1H4, #5869, Cell Signaling Technology), rabbit monoclonal anti-ULK1 (D8H5, #8054, Cell Signaling Technology), anti-pBECN1 S93/S96 (in mouse S91/S94) (#12476, Cell Signaling Technology), rabbit polyclonal anti-pBECN1 S15 (#84966, Cell Signaling Technology), rabbit polyclonal anti-BECN1 (#3738, Cell Signaling Technology), rabbit polyclonal anti-TSC2 (#3612, Cell Signaling Technology), rabbit polyclonal anti-pTSC2 S1387 (#5584, Cell Signaling Technology), rabbit monoclonal anti-pATG16L1 S278 (EPR19016, ab195242, Abcam), rabbit polyclonal anti-WIPI4 (WDR45) (19194-1-AP, Proteintech), mouse monoclonal anti-WIPI4 (G12, sc-398272, Santa Cruz Biotechnology), rabbit polyclonal anti-WIPI3 (WDR45L) (SAB2102704, Sigma-Aldrich), mouse monoclonal anti-WIPI3 (B-7, sc-514194, Santa Cruz Biotechnology), rabbit polyclonal anti-WIPI2 (#8567, Cell Signaling Technology), rabbit polyclonal anti-WIPI1 (HPA007493, Sigma-Aldrich), rabbit polyclonal anti-AMPKα1 (#2795, Cell Signaling Technology), rabbit polyclonal anti-AMPKγ2 (#2536, Cell Signaling Technology), rabbit polyclonal anti-AMPKα2 (#2757, Cell Signaling Technology), rabbit monoclonal anti-AMPKβ1 (71C10, #4178, Cell Signaling Technology), rabbit polyclonal anti-AMPKγ1 (#4187, Cell Signaling Technology), rabbit polyclonal anti-AMPKβ2 (#4188, Cell Signaling Technology), rabbit polyclonal anti-AMPKγ3 (#2550, Cell Signaling Technology), rabbit monoclonal anti-TSC1 (D43E2, #6935, Cell Signaling Technology), rabbit polyclonal anti-Flag (600-401-383, Rockland Inc.), rabbit polyclonal anti-hypusine (ABS1046, Merck Millipore), rabbit monoclonal anti-eIF5A (D8L8Q, #20765, Cell Signaling Technology), and rabbit polyclonal anti-TFEB (ab245350, Abcam).

Techniques: Over Expression, Binding Assay, Phospho-proteomics, Marker, Virus, Expressing, Control

FKBP51 overexpression is depicted in blue, and FKBP51 deletion is depicted in green. ( A and B ) Representative decrease in tissue NE content after α-MPT injection (left) and turnover rate (right) were determined on SM and eWAT (see fig. S8 for pancreas, heart, iWAT, and BAT tissues). Quantification of ( C ) pAMPK (T172) and ( D ) pp70S6K (T389), and ( E ) p62 level in the SM and eWAT. ( F ) Representative blots. ( G to H ) FKBP51 overexpression increased autophagic flux and in SM and eWAT. ( I ) Representative blots of chloroquine the experiment. Quantification of ( J ) pAMPK (T172), ( K ) pp70S6K (T389), ( L ) LC3B-II, and ( M ) p62 levels in SM and eWAT in animals lacking FKBP51 in the MBH. ( N ) Representative blots of FKBP51 MBH-KO protein analysis. All data are shown as ±SEM. Protein data are shown as the relative protein expression compared to control. A two-way ANOVA was performed, followed by a Tukey’s multiple comparison test in (F) and (G). For (A) to (E) and (I) to (L), an unpaired Student’s t test was performed. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Journal: Science Advances

Article Title: Mediobasal hypothalamic FKBP51 acts as a molecular switch linking autophagy to whole-body metabolism

doi: 10.1126/sciadv.abi4797

Figure Lengend Snippet: FKBP51 overexpression is depicted in blue, and FKBP51 deletion is depicted in green. ( A and B ) Representative decrease in tissue NE content after α-MPT injection (left) and turnover rate (right) were determined on SM and eWAT (see fig. S8 for pancreas, heart, iWAT, and BAT tissues). Quantification of ( C ) pAMPK (T172) and ( D ) pp70S6K (T389), and ( E ) p62 level in the SM and eWAT. ( F ) Representative blots. ( G to H ) FKBP51 overexpression increased autophagic flux and in SM and eWAT. ( I ) Representative blots of chloroquine the experiment. Quantification of ( J ) pAMPK (T172), ( K ) pp70S6K (T389), ( L ) LC3B-II, and ( M ) p62 levels in SM and eWAT in animals lacking FKBP51 in the MBH. ( N ) Representative blots of FKBP51 MBH-KO protein analysis. All data are shown as ±SEM. Protein data are shown as the relative protein expression compared to control. A two-way ANOVA was performed, followed by a Tukey’s multiple comparison test in (F) and (G). For (A) to (E) and (I) to (L), an unpaired Student’s t test was performed. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: The following antibodies were used: goat polyclonal anti-actin (I-19) (sc-1616, Santa Cruz Biotechnology), rabbit polyclonal anti-FKBP51 (A301-430A, Bethyl Laboratories), rabbit monoclonal anti-FKBP5 (D5G2, #12210, Cell Signaling Technology), rabbit monoclonal anti-LKB1 (D60C5, #3047, Cell Signaling Technology), rabbit polyclonal anti-pAMPKα T172 (#2531, Cell Signaling Technology), rabbit polyclonal anti-pAMPKα (#2532, Cell Signaling Technology), rabbit polyclonal anti-SKP2 (L70, #4313, Cell Signaling Technology), rabbit anti-pSKP2 S72 (was a gift from Cell Signaling Technology), rabbit polyclonal anti-AKT (#9272, Cell Signaling Technology), rabbit monoclonal anti-pAKT S473 (D9E, #4060, Cell Signaling Technology), rabbit polyclonal anti-p62 (#5114, Cell Signaling Technology), rabbit monoclonal anti-LC3B (D11, #3868, Cell Signaling Technology), rabbit polyclonal anti-pULK1 S757 (#6888, Cell Signaling Technology), rabbit monoclonal anti-pULK1 S555 (D1H4, #5869, Cell Signaling Technology), rabbit monoclonal anti-ULK1 (D8H5, #8054, Cell Signaling Technology), anti-pBECN1 S93/S96 (in mouse S91/S94) (#12476, Cell Signaling Technology), rabbit polyclonal anti-pBECN1 S15 (#84966, Cell Signaling Technology), rabbit polyclonal anti-BECN1 (#3738, Cell Signaling Technology), rabbit polyclonal anti-TSC2 (#3612, Cell Signaling Technology), rabbit polyclonal anti-pTSC2 S1387 (#5584, Cell Signaling Technology), rabbit monoclonal anti-pATG16L1 S278 (EPR19016, ab195242, Abcam), rabbit polyclonal anti-WIPI4 (WDR45) (19194-1-AP, Proteintech), mouse monoclonal anti-WIPI4 (G12, sc-398272, Santa Cruz Biotechnology), rabbit polyclonal anti-WIPI3 (WDR45L) (SAB2102704, Sigma-Aldrich), mouse monoclonal anti-WIPI3 (B-7, sc-514194, Santa Cruz Biotechnology), rabbit polyclonal anti-WIPI2 (#8567, Cell Signaling Technology), rabbit polyclonal anti-WIPI1 (HPA007493, Sigma-Aldrich), rabbit polyclonal anti-AMPKα1 (#2795, Cell Signaling Technology), rabbit polyclonal anti-AMPKγ2 (#2536, Cell Signaling Technology), rabbit polyclonal anti-AMPKα2 (#2757, Cell Signaling Technology), rabbit monoclonal anti-AMPKβ1 (71C10, #4178, Cell Signaling Technology), rabbit polyclonal anti-AMPKγ1 (#4187, Cell Signaling Technology), rabbit polyclonal anti-AMPKβ2 (#4188, Cell Signaling Technology), rabbit polyclonal anti-AMPKγ3 (#2550, Cell Signaling Technology), rabbit monoclonal anti-TSC1 (D43E2, #6935, Cell Signaling Technology), rabbit polyclonal anti-Flag (600-401-383, Rockland Inc.), rabbit polyclonal anti-hypusine (ABS1046, Merck Millipore), rabbit monoclonal anti-eIF5A (D8L8Q, #20765, Cell Signaling Technology), and rabbit polyclonal anti-TFEB (ab245350, Abcam).

Techniques: Over Expression, Injection, Expressing, Control, Comparison

( A ) Representative Western blots of phosphorylated AMP-activated protein kinase (pAMPK) and total AMPK performed in TA muscles from sedentary mice and exercised mice treated with vehicle, BCAAs , mix 1 , mix 2 , or mix 3 . ( B ) Calculation of the pAMPK/AMPK ratio for each experimental group. Values are expressed as mean ± SEM from the number of mice indicated in brackets. No statistically significant differences between vehicle and treated groups were found by one-way ANOVA followed by Bonferroni post hoc correction. Individual comparisons of the means vs. sedentary mice showed statistically significant differences found by unpaired Student’s t -test, which are indicated above the bars.

Journal: Nutrients

Article Title: Ergogenic Effect of BCAAs and L-Alanine Supplementation: Proof-of-Concept Study in a Murine Model of Physiological Exercise

doi: 10.3390/nu12082295

Figure Lengend Snippet: ( A ) Representative Western blots of phosphorylated AMP-activated protein kinase (pAMPK) and total AMPK performed in TA muscles from sedentary mice and exercised mice treated with vehicle, BCAAs , mix 1 , mix 2 , or mix 3 . ( B ) Calculation of the pAMPK/AMPK ratio for each experimental group. Values are expressed as mean ± SEM from the number of mice indicated in brackets. No statistically significant differences between vehicle and treated groups were found by one-way ANOVA followed by Bonferroni post hoc correction. Individual comparisons of the means vs. sedentary mice showed statistically significant differences found by unpaired Student’s t -test, which are indicated above the bars.

Article Snippet: The following antibodies were used: AMPK primary antibody rabbit polyclonal (Cell Signaling Technology Inc., MA, USA; dilution 1:1000); phosphorylated AMPK (pAMPK) primary antibody rabbit polyclonal (Cell Signaling Technology Inc.; dilution 1:1000); secondary antibody anti-rabbit immunoglobulin G (IgG, Sigma-Aldrich, USA; dilution 1:5000).

Techniques: Western Blot, Muscles

KEY RESOURCES TABLE

Journal: Developmental cell

Article Title: Metabolic Control over mTOR-Dependent Diapause-like State

doi: 10.1016/j.devcel.2019.12.018

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: The primary antibodies used for immunostaining were anti-RUNX1 (OriGene, TA307515), anti-Oct-4 (Santa Cruz, sc-5279), anti-pAMPKa (Thr172 Cell Signaling 40H9) and anti- Lkb1 (Santa Cruz, sc-5638).

Techniques: Recombinant, Software