rabbit polyclonal antibodies targeting pampk thr172 cat Search Results


anti pampk  (Bioss)
93
Bioss anti pampk
Effect of different protein sources on the expression of hepatic peroxisome proliferator-activated <t>receptor-γ</t> <t>(PPAR-γ),</t> uncoupling protein 2 (UCP2), phosphorylated AMP-activated protein kinase <t>(pAMPK),</t> and phosphorylated acetyl CoA carboxylase (pACC) in obese mice. ( A ) Effect of different proteins on hepatic PPAR-γ and UCP2 on gene expression in obese mice; ( B ) effect of different proteins on hepatic PPAR-γ, UCP2, pAMPK, and pACC on protein expression in obese mice. Different letters indicate significant differences between groups ( p < 0.05).
Anti Pampk, supplied by Bioss, 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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rabbit polyclonal anti pampkα t172  (Cell Signaling Technology Inc)
96
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; $$ P < 0.01. Two-way ANOVA was performed in (A) to (C) and followed by a Tukey’s multiple comparisons test. One-way ANOVA was performed for (I) and (L), followed by a Dunnett’s multiple comparison test. The unpaired Student’s t test was performed for (E) to (G). *, significant genotype effect; $, significant starvation effect; #, significant treatment effect.
Rabbit Polyclonal Anti Pampkα T172, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 96 stars, based on 1 article reviews
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rabbit monoclonal pampk thr172  (Cell Signaling Technology Inc)
95
Cell Signaling Technology Inc rabbit monoclonal pampk thr172
( A ) WT or FKBP51 KO cells were starved in HBSS medium for 4 hours to induce autophagy, followed by quantification of <t>pAMPKα</t> <t>(T172),</t> ( B ) p62, and ( C ) pp70S6K (T389). Representative blots are shown in ( D ). FKBP51 overexpression (FKBP51 OE) in N2a cells (see fig. S3D for validation) enhanced autophagy signaling. Quantification of ( E ) pAMPKα (T172), ( F ) pp70S6K (T389), ( G ) p62, and ( H ) representative blots. ( I ) Quantification of autophagic flux in FKBP51 KO and FKBP51 OE cells in response to starvation. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. ( J ) Representative blots of autophagic flux measurements. ( K ) Representative pictures of TFEB nuclear localization/translocation. DAPI, 4′,6-diamidino-2-phenylindole. Scale bar, 10 μm. ( L ) Quantification of TFEB reporter assay. BL, baseline. All data (A to J) are shown as relative fold change compared to control condition; ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001; ## P < 0.01, ### P < 0.001; $$ 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 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
https://www.bioz.com/product/rabbit+polyclonal+antibodies+targeting+pampk+thr172+cat/Phospho-AMPKalpha+(Thr172)+(D79%2E5E)+Rabbit+mAb/pmc07886321-68-57-62
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anti pampk  (Cell Signaling Technology Inc)
98
Cell Signaling Technology Inc anti pampk
( 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; $$ P < 0.01. Two-way ANOVA was performed in (A) to (C) and followed by a Tukey’s multiple comparisons test. One-way ANOVA was performed for (I) and (L), followed by a Dunnett’s multiple comparison test. The unpaired Student’s t test was performed for (E) to (G). *, significant genotype effect; $, significant starvation effect; #, significant treatment effect.
Anti Pampk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti pampkα t172  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc anti pampkα t172
( A ) WT or FKBP51 KO cells were starved in HBSS medium for 4 hours to induce autophagy, followed by quantification of <t>pAMPKα</t> <t>(T172),</t> ( B ) p62, and ( C ) pp70S6K (T389). Representative blots are shown in ( D ). FKBP51 overexpression (FKBP51 OE) in N2a cells (see fig. S3D for validation) enhanced autophagy signaling. Quantification of ( E ) pAMPKα (T172), ( F ) pp70S6K (T389), ( G ) p62, and ( H ) representative blots. ( I ) Quantification of autophagic flux in FKBP51 KO and FKBP51 OE cells in response to starvation. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. ( J ) Representative blots of autophagic flux measurements. ( K ) Representative pictures of TFEB nuclear localization/translocation. DAPI, 4′,6-diamidino-2-phenylindole. Scale bar, 10 μm. ( L ) Quantification of TFEB reporter assay. BL, baseline. All data (A to J) are shown as relative fold change compared to control condition; ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001; ## P < 0.01, ### P < 0.001; $$ 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.
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
https://www.bioz.com/product/rabbit+polyclonal+antibodies+targeting+pampk+thr172+cat/Phospho-AMPKalpha+(Thr172)+Rabbit+mAb/bio_rxiv__2025__02__15__638470-141-23-50
Average 96 stars, based on 1 article reviews
anti pampkα t172 - by Bioz Stars, 2026-09
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pampk α1 2  (Santa Cruz Biotechnology)
85
Santa Cruz Biotechnology pampk α1 2
( 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; $$ P < 0.01. Two-way ANOVA was performed in (A) to (C) and followed by a Tukey’s multiple comparisons test. One-way ANOVA was performed for (I) and (L), followed by a Dunnett’s multiple comparison test. The unpaired Student’s t test was performed for (E) to (G). *, significant genotype effect; $, significant starvation effect; #, significant treatment effect.
Pampk α1 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pampk thr172  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc pampk thr172
( A ) WT or FKBP51 KO cells were starved in HBSS medium for 4 hours to induce autophagy, followed by quantification of <t>pAMPKα</t> <t>(T172),</t> ( B ) p62, and ( C ) pp70S6K (T389). Representative blots are shown in ( D ). FKBP51 overexpression (FKBP51 OE) in N2a cells (see fig. S3D for validation) enhanced autophagy signaling. Quantification of ( E ) pAMPKα (T172), ( F ) pp70S6K (T389), ( G ) p62, and ( H ) representative blots. ( I ) Quantification of autophagic flux in FKBP51 KO and FKBP51 OE cells in response to starvation. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. ( J ) Representative blots of autophagic flux measurements. ( K ) Representative pictures of TFEB nuclear localization/translocation. DAPI, 4′,6-diamidino-2-phenylindole. Scale bar, 10 μm. ( L ) Quantification of TFEB reporter assay. BL, baseline. All data (A to J) are shown as relative fold change compared to control condition; ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001; ## P < 0.01, ### P < 0.001; $$ 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.
Pampk Thr172, 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
https://www.bioz.com/product/rabbit+polyclonal+antibodies+targeting+pampk+thr172+cat/Acetyl-CoA+Carboxylase+Rabbit+mAb/pmc07290332-53-21-24
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rabbit monoclonal pampk  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc rabbit monoclonal pampk
Effect of AdipoRon on inflammatory markers and anti-oxidant mechanisms in the mouse stomach in EtOH-induced gastric ulcer model. Figure shows changes in ( A ) MPO activity, ( B ) IL-1β levels, ( C ) <t>pAMPK/AMPK</t> ratio, ( D ) GSH, ( E ) CAT, ( F ) SOD activity, and ( G ) GPX activity. AdipoRon at the dose of 50 mg/kg p.o. significantly reduced MPO activity, IL-1β expression, and CAT activity. Moreover, AdipoRon increased GSH levels and SOD activity. $ p < 0.05 and $$ p < 0.01, as compared to control mice, and * p < 0.05 and ** p < 0.01 as compared to EtOH-group. Results are expressed as mean ± SEM for n = 5–6 mice per group.
Rabbit Monoclonal Pampk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+polyclonal+antibodies+targeting+pampk+thr172+cat/GAPDH+Rabbit+mAb/pmc08156685-195-47-52
Average 96 stars, based on 1 article reviews
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antibodies rabbit anti pampkα thr172  (Cell Signaling Technology Inc)
94
Cell Signaling Technology Inc antibodies rabbit anti pampkα thr172
Effect of AdipoRon on inflammatory markers and anti-oxidant mechanisms in the mouse stomach in EtOH-induced gastric ulcer model. Figure shows changes in ( A ) MPO activity, ( B ) IL-1β levels, ( C ) <t>pAMPK/AMPK</t> ratio, ( D ) GSH, ( E ) CAT, ( F ) SOD activity, and ( G ) GPX activity. AdipoRon at the dose of 50 mg/kg p.o. significantly reduced MPO activity, IL-1β expression, and CAT activity. Moreover, AdipoRon increased GSH levels and SOD activity. $ p < 0.05 and $$ p < 0.01, as compared to control mice, and * p < 0.05 and ** p < 0.01 as compared to EtOH-group. Results are expressed as mean ± SEM for n = 5–6 mice per group.
Antibodies Rabbit Anti Pampkα Thr172, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti phosphorylated ampk  (Cell Signaling Technology Inc)
94
Cell Signaling Technology Inc rabbit anti phosphorylated ampk
Fig. 1 Expression profile of MC4R and <t>p-AMPK</t> after ICH. a Representative western blot bands of time course and quantitative analyses of MC4R expression in the ipsilateral hemisphere after ICH. b Representative western blot bands of time course and quantitative analyses of phosphory- lated AMPK expression in the ipsilateral hemisphere after ICH. *p < 0.05 vs sham. Error bars are represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 6 per group. c Brain sample with schematic illustration showing the four areas in the perihematomal region (indicated by black squares) from where the images were taken for immunofluorescence staining (pictures are shown in panel d). d Representative images of co- localization of MC4R (red) with microglia/macrophage (Iba-1, green), neurons (NeuN, green), and astrocytes (GFAP, green) in the perihematomal area at 24 h after ICH. Nuclei were stained with DAPI (blue). Scale bar = 50 μm, n = 2
Rabbit Anti Phosphorylated Ampk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti phosphorylated ampk α1 2  (Santa Cruz Biotechnology)
94
Santa Cruz Biotechnology rabbit anti phosphorylated ampk α1 2
Fig. 1 Expression profile of MC4R and <t>p-AMPK</t> after ICH. a Representative western blot bands of time course and quantitative analyses of MC4R expression in the ipsilateral hemisphere after ICH. b Representative western blot bands of time course and quantitative analyses of phosphory- lated AMPK expression in the ipsilateral hemisphere after ICH. *p < 0.05 vs sham. Error bars are represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 6 per group. c Brain sample with schematic illustration showing the four areas in the perihematomal region (indicated by black squares) from where the images were taken for immunofluorescence staining (pictures are shown in panel d). d Representative images of co- localization of MC4R (red) with microglia/macrophage (Iba-1, green), neurons (NeuN, green), and astrocytes (GFAP, green) in the perihematomal area at 24 h after ICH. Nuclei were stained with DAPI (blue). Scale bar = 50 μm, n = 2
Rabbit Anti Phosphorylated Ampk α1 2, supplied by Santa Cruz Biotechnology, 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/product/rabbit+polyclonal+antibodies+targeting+pampk+thr172+cat/AMPK%CE%B11+Antibody/pmc05960832-63-15-18
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pampk  (Danaher Inc)
99
Danaher Inc pampk
Fig. 1 Expression profile of MC4R and <t>p-AMPK</t> after ICH. a Representative western blot bands of time course and quantitative analyses of MC4R expression in the ipsilateral hemisphere after ICH. b Representative western blot bands of time course and quantitative analyses of phosphory- lated AMPK expression in the ipsilateral hemisphere after ICH. *p < 0.05 vs sham. Error bars are represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 6 per group. c Brain sample with schematic illustration showing the four areas in the perihematomal region (indicated by black squares) from where the images were taken for immunofluorescence staining (pictures are shown in panel d). d Representative images of co- localization of MC4R (red) with microglia/macrophage (Iba-1, green), neurons (NeuN, green), and astrocytes (GFAP, green) in the perihematomal area at 24 h after ICH. Nuclei were stained with DAPI (blue). Scale bar = 50 μm, n = 2
Pampk, supplied by Danaher 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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Image Search Results


Effect of different protein sources on the expression of hepatic peroxisome proliferator-activated receptor-γ (PPAR-γ), uncoupling protein 2 (UCP2), phosphorylated AMP-activated protein kinase (pAMPK), and phosphorylated acetyl CoA carboxylase (pACC) in obese mice. ( A ) Effect of different proteins on hepatic PPAR-γ and UCP2 on gene expression in obese mice; ( B ) effect of different proteins on hepatic PPAR-γ, UCP2, pAMPK, and pACC on protein expression in obese mice. Different letters indicate significant differences between groups ( p < 0.05).

Journal: Foods

Article Title: A Comparison Study on the Therapeutic Effect of High Protein Diets Based on Pork Protein versus Soybean Protein on Obese Mice

doi: 10.3390/foods11091227

Figure Lengend Snippet: Effect of different protein sources on the expression of hepatic peroxisome proliferator-activated receptor-γ (PPAR-γ), uncoupling protein 2 (UCP2), phosphorylated AMP-activated protein kinase (pAMPK), and phosphorylated acetyl CoA carboxylase (pACC) in obese mice. ( A ) Effect of different proteins on hepatic PPAR-γ and UCP2 on gene expression in obese mice; ( B ) effect of different proteins on hepatic PPAR-γ, UCP2, pAMPK, and pACC on protein expression in obese mice. Different letters indicate significant differences between groups ( p < 0.05).

Article Snippet: The membranes were blocked in 5% BSA prepared in 1× Tris-buffered saline with 20% Tween 20 (TBST) for 1 h. The membranes were incubated overnight with anti-PPAR-γ (bs-0530R, 1:1000), anti-UCP2 (bs-1926R, 1:1000), anti-pAMPK (bs-4002R, 1:1000), anti-pACC (bs-3039R, 1:1000), anti-AMPK (bs-2771R, 1:1000) or anti-β-actin (bs-10966R, 1:1000) antibody (Bioss, Beijing, China).

Techniques: Expressing

( 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

Effect of AdipoRon on inflammatory markers and anti-oxidant mechanisms in the mouse stomach in EtOH-induced gastric ulcer model. Figure shows changes in ( A ) MPO activity, ( B ) IL-1β levels, ( C ) pAMPK/AMPK ratio, ( D ) GSH, ( E ) CAT, ( F ) SOD activity, and ( G ) GPX activity. AdipoRon at the dose of 50 mg/kg p.o. significantly reduced MPO activity, IL-1β expression, and CAT activity. Moreover, AdipoRon increased GSH levels and SOD activity. $ p < 0.05 and $$ p < 0.01, as compared to control mice, and * p < 0.05 and ** p < 0.01 as compared to EtOH-group. Results are expressed as mean ± SEM for n = 5–6 mice per group.

Journal: Molecules

Article Title: AdipoRon, an Orally Active, Synthetic Agonist of AdipoR1 and AdipoR2 Receptors Has Gastroprotective Effect in Experimentally Induced Gastric Ulcers in Mice

doi: 10.3390/molecules26102946

Figure Lengend Snippet: Effect of AdipoRon on inflammatory markers and anti-oxidant mechanisms in the mouse stomach in EtOH-induced gastric ulcer model. Figure shows changes in ( A ) MPO activity, ( B ) IL-1β levels, ( C ) pAMPK/AMPK ratio, ( D ) GSH, ( E ) CAT, ( F ) SOD activity, and ( G ) GPX activity. AdipoRon at the dose of 50 mg/kg p.o. significantly reduced MPO activity, IL-1β expression, and CAT activity. Moreover, AdipoRon increased GSH levels and SOD activity. $ p < 0.05 and $$ p < 0.01, as compared to control mice, and * p < 0.05 and ** p < 0.01 as compared to EtOH-group. Results are expressed as mean ± SEM for n = 5–6 mice per group.

Article Snippet: Then, membranes were probed with the following primary antibodies diluted in 1% non-fat dry milk in PBST for 80 min at 25 °C: mouse monoclonal IL-1β (1:1000, sc-32294 1:1000 Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit monoclonal AMPK antibody (1:1000, D5A2, Cell Signaling Technology, Massachusetts, USA); rabbit monoclonal pAMPK (1:1000, Thr172, Cell Signaling Technology, Massachusetts, USA), glyceraldehyde-3-phosphate dehydrogenase (GAPDH; 1:15 000; MAB374; Merck Millipore, Warsaw, Poland) as a reference protein.

Techniques: Activity Assay, Expressing, Control

Effect of AdipoRon on inflammatory markers and antioxidant mechanisms in the mouse stomach in diclofenac-induced gastric ulcer model. Figure shows changes in ( A ) MPO activity, ( B ) IL-1β levels, ( C ) pAMPK/AMPK ratio, ( D ) GSH, ( E ) catalase, ( F ) SOD, and ( G ) GPX activity. AdipoRon 50 mg/kg p.o. reduced MPO activity, IL-1β expression, and SOD activity and increased GPX activity. AdipoRon had no effect on GSH levels and CAT activity in diclofenac induced model of gastric ulcer in mice. $ p < 0.05, as compared to control mice, and * p < 0.05 as compared to diclofenac-group. Results are expressed as mean ± SEM for n = 5–6 mice per group.

Journal: Molecules

Article Title: AdipoRon, an Orally Active, Synthetic Agonist of AdipoR1 and AdipoR2 Receptors Has Gastroprotective Effect in Experimentally Induced Gastric Ulcers in Mice

doi: 10.3390/molecules26102946

Figure Lengend Snippet: Effect of AdipoRon on inflammatory markers and antioxidant mechanisms in the mouse stomach in diclofenac-induced gastric ulcer model. Figure shows changes in ( A ) MPO activity, ( B ) IL-1β levels, ( C ) pAMPK/AMPK ratio, ( D ) GSH, ( E ) catalase, ( F ) SOD, and ( G ) GPX activity. AdipoRon 50 mg/kg p.o. reduced MPO activity, IL-1β expression, and SOD activity and increased GPX activity. AdipoRon had no effect on GSH levels and CAT activity in diclofenac induced model of gastric ulcer in mice. $ p < 0.05, as compared to control mice, and * p < 0.05 as compared to diclofenac-group. Results are expressed as mean ± SEM for n = 5–6 mice per group.

Article Snippet: Then, membranes were probed with the following primary antibodies diluted in 1% non-fat dry milk in PBST for 80 min at 25 °C: mouse monoclonal IL-1β (1:1000, sc-32294 1:1000 Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit monoclonal AMPK antibody (1:1000, D5A2, Cell Signaling Technology, Massachusetts, USA); rabbit monoclonal pAMPK (1:1000, Thr172, Cell Signaling Technology, Massachusetts, USA), glyceraldehyde-3-phosphate dehydrogenase (GAPDH; 1:15 000; MAB374; Merck Millipore, Warsaw, Poland) as a reference protein.

Techniques: Activity Assay, Expressing, Control

Fig. 1 Expression profile of MC4R and p-AMPK after ICH. a Representative western blot bands of time course and quantitative analyses of MC4R expression in the ipsilateral hemisphere after ICH. b Representative western blot bands of time course and quantitative analyses of phosphory- lated AMPK expression in the ipsilateral hemisphere after ICH. *p < 0.05 vs sham. Error bars are represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 6 per group. c Brain sample with schematic illustration showing the four areas in the perihematomal region (indicated by black squares) from where the images were taken for immunofluorescence staining (pictures are shown in panel d). d Representative images of co- localization of MC4R (red) with microglia/macrophage (Iba-1, green), neurons (NeuN, green), and astrocytes (GFAP, green) in the perihematomal area at 24 h after ICH. Nuclei were stained with DAPI (blue). Scale bar = 50 μm, n = 2

Journal: Journal of neuroinflammation

Article Title: Activation of melanocortin receptor 4 with RO27-3225 attenuates neuroinflammation through AMPK/JNK/p38 MAPK pathway after intracerebral hemorrhage in mice.

doi: 10.1186/s12974-018-1140-6

Figure Lengend Snippet: Fig. 1 Expression profile of MC4R and p-AMPK after ICH. a Representative western blot bands of time course and quantitative analyses of MC4R expression in the ipsilateral hemisphere after ICH. b Representative western blot bands of time course and quantitative analyses of phosphory- lated AMPK expression in the ipsilateral hemisphere after ICH. *p < 0.05 vs sham. Error bars are represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 6 per group. c Brain sample with schematic illustration showing the four areas in the perihematomal region (indicated by black squares) from where the images were taken for immunofluorescence staining (pictures are shown in panel d). d Representative images of co- localization of MC4R (red) with microglia/macrophage (Iba-1, green), neurons (NeuN, green), and astrocytes (GFAP, green) in the perihematomal area at 24 h after ICH. Nuclei were stained with DAPI (blue). Scale bar = 50 μm, n = 2

Article Snippet: The membrane was blocked and incubated at 4 °C overnight with the following primary antibodies: rabbit anti-MC4R (1:500, Abcam, Cambridge, MA), rabbit anti-AMPK (1:1000, Cell Signaling, Danvers, MA), rabbit anti-phosphorylated AMPK (p-AMPK, Thr 172, 1:1000, Cell Signaling), rabbit anti-JNK (1:1000, Abcam), rabbit anti-phosphorylated JNK (p-JNK, 1:1000, Abcam), mouse anti-p38 (1:300, Santa Cruz Biotechnology, Santa Cruz, CA), mouse anti-phosphorylated p38 (p-p38, 1:300, Santa Cruz), goat anti-Iba-1 (1:1000, Abcam), rabbit anti-MPO (1:500, Abcam), rabbit anti-TNF-α (1:1000, Abcam), rabbit anti-IL-1β (1:1000, Abcam), and goat antiactin (1:5000, Santa Cruz).

Techniques: Expressing, Western Blot, Immunofluorescence, Staining

Fig. 5 MC4R antagonist and AMPK inhibitor reversed the effects of RO27-3225 on neurobehavioral outcomes after ICH. a Modified Garcia test, b forelimb placement test, c corner turn test, and d the effects of RO27-3225 on body weight loss at 24 h after ICH. *p < 0.05 vs. sham, #p < 0.05 vs. ICH + vehicle, @p < 0.05 vs. ICH + RO27-3225 + HS024, and &p < 0.05 vs. ICH + RO27-3225 + DMSO. Error bars are represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 6 per group

Journal: Journal of neuroinflammation

Article Title: Activation of melanocortin receptor 4 with RO27-3225 attenuates neuroinflammation through AMPK/JNK/p38 MAPK pathway after intracerebral hemorrhage in mice.

doi: 10.1186/s12974-018-1140-6

Figure Lengend Snippet: Fig. 5 MC4R antagonist and AMPK inhibitor reversed the effects of RO27-3225 on neurobehavioral outcomes after ICH. a Modified Garcia test, b forelimb placement test, c corner turn test, and d the effects of RO27-3225 on body weight loss at 24 h after ICH. *p < 0.05 vs. sham, #p < 0.05 vs. ICH + vehicle, @p < 0.05 vs. ICH + RO27-3225 + HS024, and &p < 0.05 vs. ICH + RO27-3225 + DMSO. Error bars are represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 6 per group

Article Snippet: The membrane was blocked and incubated at 4 °C overnight with the following primary antibodies: rabbit anti-MC4R (1:500, Abcam, Cambridge, MA), rabbit anti-AMPK (1:1000, Cell Signaling, Danvers, MA), rabbit anti-phosphorylated AMPK (p-AMPK, Thr 172, 1:1000, Cell Signaling), rabbit anti-JNK (1:1000, Abcam), rabbit anti-phosphorylated JNK (p-JNK, 1:1000, Abcam), mouse anti-p38 (1:300, Santa Cruz Biotechnology, Santa Cruz, CA), mouse anti-phosphorylated p38 (p-p38, 1:300, Santa Cruz), goat anti-Iba-1 (1:1000, Abcam), rabbit anti-MPO (1:500, Abcam), rabbit anti-TNF-α (1:1000, Abcam), rabbit anti-IL-1β (1:1000, Abcam), and goat antiactin (1:5000, Santa Cruz).

Techniques: Modification

Fig. 6 The effects of RO27-3225 and MC4R antagonist HS024 on the expression of MC4R and its downstream signaling proteins. a Representative western blot bands. b–g Quantitative analyses of MC4R, phosphorylated AMPK, phosphorylated JNK, phosphorylated p38 MAPK, TNF-α, and IL-1β in the ipsilateral hemisphere at 24 h after ICH. *p < 0.05 vs. sham, #p < 0.05 vs. ICH + vehicle, and @p < 0.05 vs. ICH + RO27-3225 + HS024. Error bars are represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 6 per group

Journal: Journal of neuroinflammation

Article Title: Activation of melanocortin receptor 4 with RO27-3225 attenuates neuroinflammation through AMPK/JNK/p38 MAPK pathway after intracerebral hemorrhage in mice.

doi: 10.1186/s12974-018-1140-6

Figure Lengend Snippet: Fig. 6 The effects of RO27-3225 and MC4R antagonist HS024 on the expression of MC4R and its downstream signaling proteins. a Representative western blot bands. b–g Quantitative analyses of MC4R, phosphorylated AMPK, phosphorylated JNK, phosphorylated p38 MAPK, TNF-α, and IL-1β in the ipsilateral hemisphere at 24 h after ICH. *p < 0.05 vs. sham, #p < 0.05 vs. ICH + vehicle, and @p < 0.05 vs. ICH + RO27-3225 + HS024. Error bars are represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 6 per group

Article Snippet: The membrane was blocked and incubated at 4 °C overnight with the following primary antibodies: rabbit anti-MC4R (1:500, Abcam, Cambridge, MA), rabbit anti-AMPK (1:1000, Cell Signaling, Danvers, MA), rabbit anti-phosphorylated AMPK (p-AMPK, Thr 172, 1:1000, Cell Signaling), rabbit anti-JNK (1:1000, Abcam), rabbit anti-phosphorylated JNK (p-JNK, 1:1000, Abcam), mouse anti-p38 (1:300, Santa Cruz Biotechnology, Santa Cruz, CA), mouse anti-phosphorylated p38 (p-p38, 1:300, Santa Cruz), goat anti-Iba-1 (1:1000, Abcam), rabbit anti-MPO (1:500, Abcam), rabbit anti-TNF-α (1:1000, Abcam), rabbit anti-IL-1β (1:1000, Abcam), and goat antiactin (1:5000, Santa Cruz).

Techniques: Expressing, Western Blot

Fig. 7 The effects of RO27-3225 and AMPK inhibitor dorsomorphin on the expression of MC4R and its downstream signaling proteins. a Repre- sentative western blot bands. b–g Quantitative analyses of MC4R, phosphorylated AMPK, phosphorylated JNK, phosphorylated p38 MAPK, TNF-α, and IL-1β in the ipsilateral hemisphere at 24 h after ICH. *p < 0.05 vs. sham, #p < 0.05 vs. ICH + vehicle, and &p < 0.05 vs. ICH + RO27-3225 + DMSO. Error bars are represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 6 per group

Journal: Journal of neuroinflammation

Article Title: Activation of melanocortin receptor 4 with RO27-3225 attenuates neuroinflammation through AMPK/JNK/p38 MAPK pathway after intracerebral hemorrhage in mice.

doi: 10.1186/s12974-018-1140-6

Figure Lengend Snippet: Fig. 7 The effects of RO27-3225 and AMPK inhibitor dorsomorphin on the expression of MC4R and its downstream signaling proteins. a Repre- sentative western blot bands. b–g Quantitative analyses of MC4R, phosphorylated AMPK, phosphorylated JNK, phosphorylated p38 MAPK, TNF-α, and IL-1β in the ipsilateral hemisphere at 24 h after ICH. *p < 0.05 vs. sham, #p < 0.05 vs. ICH + vehicle, and &p < 0.05 vs. ICH + RO27-3225 + DMSO. Error bars are represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 6 per group

Article Snippet: The membrane was blocked and incubated at 4 °C overnight with the following primary antibodies: rabbit anti-MC4R (1:500, Abcam, Cambridge, MA), rabbit anti-AMPK (1:1000, Cell Signaling, Danvers, MA), rabbit anti-phosphorylated AMPK (p-AMPK, Thr 172, 1:1000, Cell Signaling), rabbit anti-JNK (1:1000, Abcam), rabbit anti-phosphorylated JNK (p-JNK, 1:1000, Abcam), mouse anti-p38 (1:300, Santa Cruz Biotechnology, Santa Cruz, CA), mouse anti-phosphorylated p38 (p-p38, 1:300, Santa Cruz), goat anti-Iba-1 (1:1000, Abcam), rabbit anti-MPO (1:500, Abcam), rabbit anti-TNF-α (1:1000, Abcam), rabbit anti-IL-1β (1:1000, Abcam), and goat antiactin (1:5000, Santa Cruz).

Techniques: Expressing, Western Blot