pampk rabbit antibody Search Results


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; $$ P < 0.01. Two-way ANOVA was performed in (A) to (C) and followed by a Tukey’s multiple comparisons test. One-way ANOVA was performed for (I) and (L), followed by a Dunnett’s multiple comparison test. The unpaired Student’s t test was performed for (E) to (G). *, significant genotype effect; $, significant starvation effect; #, significant treatment effect.
Rabbit Polyclonal Anti Pampkα T172, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pampk+rabbit+antibody/Phospho-AMPKalpha+(Thr172)+Antibody/pmc08906734-206-35-40
Average 96 stars, based on 1 article reviews
rabbit polyclonal anti pampkα t172 - by Bioz Stars, 2026-09
96/100 stars
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rabbit monoclonal pampk thr172  (Cell Signaling Technology Inc)
95
Cell Signaling Technology Inc rabbit monoclonal pampk thr172
( A ) WT or FKBP51 KO cells were starved in HBSS medium for 4 hours to induce autophagy, followed by quantification of <t>pAMPKα</t> <t>(T172),</t> ( B ) p62, and ( C ) pp70S6K (T389). Representative blots are shown in ( D ). FKBP51 overexpression (FKBP51 OE) in N2a cells (see fig. S3D for validation) enhanced autophagy signaling. Quantification of ( E ) pAMPKα (T172), ( F ) pp70S6K (T389), ( G ) p62, and ( H ) representative blots. ( I ) Quantification of autophagic flux in FKBP51 KO and FKBP51 OE cells in response to starvation. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. ( J ) Representative blots of autophagic flux measurements. ( K ) Representative pictures of TFEB nuclear localization/translocation. DAPI, 4′,6-diamidino-2-phenylindole. Scale bar, 10 μm. ( L ) Quantification of TFEB reporter assay. BL, baseline. All data (A to J) are shown as relative fold change compared to control condition; ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001; ## P < 0.01, ### P < 0.001; $$ 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/pampk+rabbit+antibody/Phospho-AMPKalpha+(Thr172)+(D79%2E5E)+Rabbit+mAb/pmc07886321-68-57-62
Average 95 stars, based on 1 article reviews
rabbit monoclonal pampk thr172 - by Bioz Stars, 2026-09
95/100 stars
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rabbit anti phosphorylated ampk p ampk  (Cell Signaling Technology Inc)
94
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; $$ P < 0.01. Two-way ANOVA was performed in (A) to (C) and followed by a Tukey’s multiple comparisons test. One-way ANOVA was performed for (I) and (L), followed by a Dunnett’s multiple comparison test. The unpaired Student’s t test was performed for (E) to (G). *, significant genotype effect; $, significant starvation effect; #, significant treatment effect.
Rabbit Anti Phosphorylated Ampk P Ampk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pampk+rabbit+antibody/Phospho-HDAC4+(Ser632)%2FHDAC5+(Ser661)%2FHDAC7+(Ser486)+Antibody/pmc10676800-59-41-47
Average 94 stars, based on 1 article reviews
rabbit anti phosphorylated ampk p ampk - by Bioz Stars, 2026-09
94/100 stars
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anti pampk  (Cell Signaling Technology Inc)
98
Cell Signaling Technology Inc anti pampk
( A ) WT or FKBP51 KO cells were starved in HBSS medium for 4 hours to induce autophagy, followed by quantification of <t>pAMPKα</t> <t>(T172),</t> ( B ) p62, and ( C ) pp70S6K (T389). Representative blots are shown in ( D ). FKBP51 overexpression (FKBP51 OE) in N2a cells (see fig. S3D for validation) enhanced autophagy signaling. Quantification of ( E ) pAMPKα (T172), ( F ) pp70S6K (T389), ( G ) p62, and ( H ) representative blots. ( I ) Quantification of autophagic flux in FKBP51 KO and FKBP51 OE cells in response to starvation. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. ( J ) Representative blots of autophagic flux measurements. ( K ) Representative pictures of TFEB nuclear localization/translocation. DAPI, 4′,6-diamidino-2-phenylindole. Scale bar, 10 μm. ( L ) Quantification of TFEB reporter assay. BL, baseline. All data (A to J) are shown as relative fold change compared to control condition; ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001; ## P < 0.01, ### P < 0.001; $$ 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
https://www.bioz.com/product/pampk+rabbit+antibody/Phospho-AMPKalpha+(Thr172)+Rabbit+mAb/pmc11425072-26-16-19
Average 98 stars, based on 1 article reviews
anti pampk - by Bioz Stars, 2026-09
98/100 stars
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rabbit anti yme1l1 polyclonal  (Proteintech)
95
Proteintech rabbit anti yme1l1 polyclonal
( 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; $$ P < 0.01. Two-way ANOVA was performed in (A) to (C) and followed by a Tukey’s multiple comparisons test. One-way ANOVA was performed for (I) and (L), followed by a Dunnett’s multiple comparison test. The unpaired Student’s t test was performed for (E) to (G). *, significant genotype effect; $, significant starvation effect; #, significant treatment effect.
Rabbit Anti Yme1l1 Polyclonal, supplied by Proteintech, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pampk+rabbit+antibody/YME1L1+Antibody/pmc09598227-63-48-53
Average 95 stars, based on 1 article reviews
rabbit anti yme1l1 polyclonal - by Bioz Stars, 2026-09
95/100 stars
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pampk antibody  (Toyobo)
98
Toyobo pampk antibody
( A ) WT or FKBP51 KO cells were starved in HBSS medium for 4 hours to induce autophagy, followed by quantification of <t>pAMPKα</t> <t>(T172),</t> ( B ) p62, and ( C ) pp70S6K (T389). Representative blots are shown in ( D ). FKBP51 overexpression (FKBP51 OE) in N2a cells (see fig. S3D for validation) enhanced autophagy signaling. Quantification of ( E ) pAMPKα (T172), ( F ) pp70S6K (T389), ( G ) p62, and ( H ) representative blots. ( I ) Quantification of autophagic flux in FKBP51 KO and FKBP51 OE cells in response to starvation. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. ( J ) Representative blots of autophagic flux measurements. ( K ) Representative pictures of TFEB nuclear localization/translocation. DAPI, 4′,6-diamidino-2-phenylindole. Scale bar, 10 μm. ( L ) Quantification of TFEB reporter assay. BL, baseline. All data (A to J) are shown as relative fold change compared to control condition; ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001; ## P < 0.01, ### P < 0.001; $$ 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.
Pampk Antibody, supplied by Toyobo, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pampk+rabbit+antibody/Can+Get+Signal/us12357626-415-2-14
Average 98 stars, based on 1 article reviews
pampk antibody - by Bioz Stars, 2026-09
98/100 stars
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rabbit anti human adm polyclonal antibody  (Boster Bio)
90
Boster Bio rabbit anti human adm polyclonal 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; $$ P < 0.01. Two-way ANOVA was performed in (A) to (C) and followed by a Tukey’s multiple comparisons test. One-way ANOVA was performed for (I) and (L), followed by a Dunnett’s multiple comparison test. The unpaired Student’s t test was performed for (E) to (G). *, significant genotype effect; $, significant starvation effect; #, significant treatment effect.
Rabbit Anti Human Adm Polyclonal Antibody, supplied by Boster Bio, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pampk+rabbit+antibody/Anti-ADM+Antibody/pmc04508992-99-1-12
Average 90 stars, based on 1 article reviews
rabbit anti human adm polyclonal antibody - by Bioz Stars, 2026-09
90/100 stars
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anti pampkα t172  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc anti pampkα t172
( A ) WT or FKBP51 KO cells were starved in HBSS medium for 4 hours to induce autophagy, followed by quantification of <t>pAMPKα</t> <t>(T172),</t> ( B ) p62, and ( C ) pp70S6K (T389). Representative blots are shown in ( D ). FKBP51 overexpression (FKBP51 OE) in N2a cells (see fig. S3D for validation) enhanced autophagy signaling. Quantification of ( E ) pAMPKα (T172), ( F ) pp70S6K (T389), ( G ) p62, and ( H ) representative blots. ( I ) Quantification of autophagic flux in FKBP51 KO and FKBP51 OE cells in response to starvation. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. ( J ) Representative blots of autophagic flux measurements. ( K ) Representative pictures of TFEB nuclear localization/translocation. DAPI, 4′,6-diamidino-2-phenylindole. Scale bar, 10 μm. ( L ) Quantification of TFEB reporter assay. BL, baseline. All data (A to J) are shown as relative fold change compared to control condition; ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001; ## P < 0.01, ### P < 0.001; $$ 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/pampk+rabbit+antibody/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
96/100 stars
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rabbit rabbit pampk  (Cell Signaling Technology Inc)
97
Cell Signaling Technology Inc rabbit rabbit 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.
Rabbit Rabbit Pampk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pampk+rabbit+antibody/AMPKalpha+Antibody/pm37892450-77-30-34
Average 97 stars, based on 1 article reviews
rabbit rabbit pampk - by Bioz Stars, 2026-09
97/100 stars
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pampk  (Cell Signaling Technology Inc)
95
Cell Signaling Technology Inc pampk
( A ) Treatment with IL6 (20 ng/mL) for 2 hours (left panel) or the induction of cancer stem cells (CSCs) (right panel) upregulates the expression of most glycolysis-related genes, including PFKL, at the mRNA level. Red bars represent PLC5 cells after IL6 treatment or sphere formation, while black bars represent control cells. ( B ) IL6 treatment increases PFKL protein levels. The right panel shows the quantification results of western blot analysis for PFKL and RPIA protein levels. Black bars indicate no treatment control, red bars represent 0.5 hours of IL6 treatment, and blue bars represent 2 hours of IL6 treatment. ( C ) Knockdown <t>of</t> <t>AMPK</t> by shRNA reduces IL6-induced PFKL-upregulated protein expression in PLC5 cells, leading to decreases in <t>pAMPK,</t> PFKL, RPIA, pERK, and pSMAD5 levels, while it has no effect on STAT3 phosphorylation. The right panel illustrates the quantification of western blot results for AMPK, pAMPK, PFKL, RPIA, STAT3, pSTAT3, ERK, pERK, mTOR, p-mTOR, SMAD5, and pSMAD5. Black bars represent no treatment control, red bars indicate IL6 treatment, blue bars represent shAMPK, and the green bar denotes shAMPK+IL6. ( D ) Knockdown of AMPK decreases cell viability in PLC5 cells. Quantification of cell viability at 24, 48, and 72 hours is shown. Red bars represent shAMPK, while black bars denote shLuc control. ( E ) AMPK knockdown reduces IL6-stimulated cell viability, normalized to the control without IL6 treatment. Black bars represent the control without treatment, and red bars represent IL6 treatment. ( F ) Inhibition of AMPK with dorsomorphin decreases cell viability in three hepatoma cell lines. Cell death rates were quantified, and the IC50 for dorsomorphin in PLC5, Hep3B, and HepG2 cells is displayed in the upper left. ( G ) AMPK knockdown decreases migration ability with or without IL6 treatment. Quantification of migration without IL6 or with IL6 treatment is shown. Black bars represent sh-Luc control, and red bars denote sh-AMPK. ( H ) Suppression of AMPK with dorsomorphin reduces migration ability. Black bars represent the control without treatment, and red bars indicate dorsomorphin treatment. ( I ) Dorsomorphin significantly reduces PFKL protein levels. The left panel presents quantification of western blot results for AMPK, pAMPK, PFKL, SMAD5, pSMAD5, ACC, and pACC. Black bars represent the control without treatment, light red bars denote 1 µg/mL, and red bars indicate 6 µg/mL dorsomorphin treatment. ( J ) AMPK knockdown does not affect PFKL mRNA expression. Quantification of qPCR results for PFKL mRNA is shown. The red bar indicates dorsomorphin treatment, while the black bar denotes the control without treatment. ( K ) AMPK knockdown increases proteasome activity in PLC5 cells, suggesting that AMPK stabilizes PFKL by inhibiting proteasome activity. Quantification of proteasome activity is shown, with the red bar indicating shAMPK and the black bar representing shLuc control. Statistical analyses were performed using one-way ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.
Pampk, 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/pampk+rabbit+antibody/Phospho-AMPK+Substrate+Motif+%5BLXRXX(pS%2FpT)+MultiMab+Rabbit+mAb+mix/bio_rxiv__2024__02__29__582877-222-32-33
Average 95 stars, based on 1 article reviews
pampk - by Bioz Stars, 2026-09
95/100 stars
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adm  (Proteintech)
93
Proteintech adm
( A ) Treatment with IL6 (20 ng/mL) for 2 hours (left panel) or the induction of cancer stem cells (CSCs) (right panel) upregulates the expression of most glycolysis-related genes, including PFKL, at the mRNA level. Red bars represent PLC5 cells after IL6 treatment or sphere formation, while black bars represent control cells. ( B ) IL6 treatment increases PFKL protein levels. The right panel shows the quantification results of western blot analysis for PFKL and RPIA protein levels. Black bars indicate no treatment control, red bars represent 0.5 hours of IL6 treatment, and blue bars represent 2 hours of IL6 treatment. ( C ) Knockdown <t>of</t> <t>AMPK</t> by shRNA reduces IL6-induced PFKL-upregulated protein expression in PLC5 cells, leading to decreases in <t>pAMPK,</t> PFKL, RPIA, pERK, and pSMAD5 levels, while it has no effect on STAT3 phosphorylation. The right panel illustrates the quantification of western blot results for AMPK, pAMPK, PFKL, RPIA, STAT3, pSTAT3, ERK, pERK, mTOR, p-mTOR, SMAD5, and pSMAD5. Black bars represent no treatment control, red bars indicate IL6 treatment, blue bars represent shAMPK, and the green bar denotes shAMPK+IL6. ( D ) Knockdown of AMPK decreases cell viability in PLC5 cells. Quantification of cell viability at 24, 48, and 72 hours is shown. Red bars represent shAMPK, while black bars denote shLuc control. ( E ) AMPK knockdown reduces IL6-stimulated cell viability, normalized to the control without IL6 treatment. Black bars represent the control without treatment, and red bars represent IL6 treatment. ( F ) Inhibition of AMPK with dorsomorphin decreases cell viability in three hepatoma cell lines. Cell death rates were quantified, and the IC50 for dorsomorphin in PLC5, Hep3B, and HepG2 cells is displayed in the upper left. ( G ) AMPK knockdown decreases migration ability with or without IL6 treatment. Quantification of migration without IL6 or with IL6 treatment is shown. Black bars represent sh-Luc control, and red bars denote sh-AMPK. ( H ) Suppression of AMPK with dorsomorphin reduces migration ability. Black bars represent the control without treatment, and red bars indicate dorsomorphin treatment. ( I ) Dorsomorphin significantly reduces PFKL protein levels. The left panel presents quantification of western blot results for AMPK, pAMPK, PFKL, SMAD5, pSMAD5, ACC, and pACC. Black bars represent the control without treatment, light red bars denote 1 µg/mL, and red bars indicate 6 µg/mL dorsomorphin treatment. ( J ) AMPK knockdown does not affect PFKL mRNA expression. Quantification of qPCR results for PFKL mRNA is shown. The red bar indicates dorsomorphin treatment, while the black bar denotes the control without treatment. ( K ) AMPK knockdown increases proteasome activity in PLC5 cells, suggesting that AMPK stabilizes PFKL by inhibiting proteasome activity. Quantification of proteasome activity is shown, with the red bar indicating shAMPK and the black bar representing shLuc control. Statistical analyses were performed using one-way ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.
Adm, supplied by Proteintech, 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/product/pampk+rabbit+antibody/Adrenomedullin+Antibody/ppr0925375-79-6-7
Average 93 stars, based on 1 article reviews
adm - by Bioz Stars, 2026-09
93/100 stars
  Buy from Supplier
rabbit polyclonal anti creb  (New England Biolabs)
96
New England Biolabs rabbit polyclonal anti creb
( A ) Treatment with IL6 (20 ng/mL) for 2 hours (left panel) or the induction of cancer stem cells (CSCs) (right panel) upregulates the expression of most glycolysis-related genes, including PFKL, at the mRNA level. Red bars represent PLC5 cells after IL6 treatment or sphere formation, while black bars represent control cells. ( B ) IL6 treatment increases PFKL protein levels. The right panel shows the quantification results of western blot analysis for PFKL and RPIA protein levels. Black bars indicate no treatment control, red bars represent 0.5 hours of IL6 treatment, and blue bars represent 2 hours of IL6 treatment. ( C ) Knockdown <t>of</t> <t>AMPK</t> by shRNA reduces IL6-induced PFKL-upregulated protein expression in PLC5 cells, leading to decreases in <t>pAMPK,</t> PFKL, RPIA, pERK, and pSMAD5 levels, while it has no effect on STAT3 phosphorylation. The right panel illustrates the quantification of western blot results for AMPK, pAMPK, PFKL, RPIA, STAT3, pSTAT3, ERK, pERK, mTOR, p-mTOR, SMAD5, and pSMAD5. Black bars represent no treatment control, red bars indicate IL6 treatment, blue bars represent shAMPK, and the green bar denotes shAMPK+IL6. ( D ) Knockdown of AMPK decreases cell viability in PLC5 cells. Quantification of cell viability at 24, 48, and 72 hours is shown. Red bars represent shAMPK, while black bars denote shLuc control. ( E ) AMPK knockdown reduces IL6-stimulated cell viability, normalized to the control without IL6 treatment. Black bars represent the control without treatment, and red bars represent IL6 treatment. ( F ) Inhibition of AMPK with dorsomorphin decreases cell viability in three hepatoma cell lines. Cell death rates were quantified, and the IC50 for dorsomorphin in PLC5, Hep3B, and HepG2 cells is displayed in the upper left. ( G ) AMPK knockdown decreases migration ability with or without IL6 treatment. Quantification of migration without IL6 or with IL6 treatment is shown. Black bars represent sh-Luc control, and red bars denote sh-AMPK. ( H ) Suppression of AMPK with dorsomorphin reduces migration ability. Black bars represent the control without treatment, and red bars indicate dorsomorphin treatment. ( I ) Dorsomorphin significantly reduces PFKL protein levels. The left panel presents quantification of western blot results for AMPK, pAMPK, PFKL, SMAD5, pSMAD5, ACC, and pACC. Black bars represent the control without treatment, light red bars denote 1 µg/mL, and red bars indicate 6 µg/mL dorsomorphin treatment. ( J ) AMPK knockdown does not affect PFKL mRNA expression. Quantification of qPCR results for PFKL mRNA is shown. The red bar indicates dorsomorphin treatment, while the black bar denotes the control without treatment. ( K ) AMPK knockdown increases proteasome activity in PLC5 cells, suggesting that AMPK stabilizes PFKL by inhibiting proteasome activity. Quantification of proteasome activity is shown, with the red bar indicating shAMPK and the black bar representing shLuc control. Statistical analyses were performed using one-way ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.
Rabbit Polyclonal Anti Creb, supplied by New England Biolabs, 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/pampk+rabbit+antibody/Anti-SNAP-tag+Antibody/pm21220102-270-22-30
Average 96 stars, based on 1 article reviews
rabbit polyclonal anti creb - by Bioz Stars, 2026-09
96/100 stars
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Image Search Results


( 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 ) Treatment with IL6 (20 ng/mL) for 2 hours (left panel) or the induction of cancer stem cells (CSCs) (right panel) upregulates the expression of most glycolysis-related genes, including PFKL, at the mRNA level. Red bars represent PLC5 cells after IL6 treatment or sphere formation, while black bars represent control cells. ( B ) IL6 treatment increases PFKL protein levels. The right panel shows the quantification results of western blot analysis for PFKL and RPIA protein levels. Black bars indicate no treatment control, red bars represent 0.5 hours of IL6 treatment, and blue bars represent 2 hours of IL6 treatment. ( C ) Knockdown of AMPK by shRNA reduces IL6-induced PFKL-upregulated protein expression in PLC5 cells, leading to decreases in pAMPK, PFKL, RPIA, pERK, and pSMAD5 levels, while it has no effect on STAT3 phosphorylation. The right panel illustrates the quantification of western blot results for AMPK, pAMPK, PFKL, RPIA, STAT3, pSTAT3, ERK, pERK, mTOR, p-mTOR, SMAD5, and pSMAD5. Black bars represent no treatment control, red bars indicate IL6 treatment, blue bars represent shAMPK, and the green bar denotes shAMPK+IL6. ( D ) Knockdown of AMPK decreases cell viability in PLC5 cells. Quantification of cell viability at 24, 48, and 72 hours is shown. Red bars represent shAMPK, while black bars denote shLuc control. ( E ) AMPK knockdown reduces IL6-stimulated cell viability, normalized to the control without IL6 treatment. Black bars represent the control without treatment, and red bars represent IL6 treatment. ( F ) Inhibition of AMPK with dorsomorphin decreases cell viability in three hepatoma cell lines. Cell death rates were quantified, and the IC50 for dorsomorphin in PLC5, Hep3B, and HepG2 cells is displayed in the upper left. ( G ) AMPK knockdown decreases migration ability with or without IL6 treatment. Quantification of migration without IL6 or with IL6 treatment is shown. Black bars represent sh-Luc control, and red bars denote sh-AMPK. ( H ) Suppression of AMPK with dorsomorphin reduces migration ability. Black bars represent the control without treatment, and red bars indicate dorsomorphin treatment. ( I ) Dorsomorphin significantly reduces PFKL protein levels. The left panel presents quantification of western blot results for AMPK, pAMPK, PFKL, SMAD5, pSMAD5, ACC, and pACC. Black bars represent the control without treatment, light red bars denote 1 µg/mL, and red bars indicate 6 µg/mL dorsomorphin treatment. ( J ) AMPK knockdown does not affect PFKL mRNA expression. Quantification of qPCR results for PFKL mRNA is shown. The red bar indicates dorsomorphin treatment, while the black bar denotes the control without treatment. ( K ) AMPK knockdown increases proteasome activity in PLC5 cells, suggesting that AMPK stabilizes PFKL by inhibiting proteasome activity. Quantification of proteasome activity is shown, with the red bar indicating shAMPK and the black bar representing shLuc control. Statistical analyses were performed using one-way ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Journal: bioRxiv

Article Title: Stabilization of AMPK/PFKL/RPIA in the Glycolytic Bodies Transduces IL6/STAT3 Signal in Hepatocarcinogenesis

doi: 10.1101/2024.02.29.582877

Figure Lengend Snippet: ( A ) Treatment with IL6 (20 ng/mL) for 2 hours (left panel) or the induction of cancer stem cells (CSCs) (right panel) upregulates the expression of most glycolysis-related genes, including PFKL, at the mRNA level. Red bars represent PLC5 cells after IL6 treatment or sphere formation, while black bars represent control cells. ( B ) IL6 treatment increases PFKL protein levels. The right panel shows the quantification results of western blot analysis for PFKL and RPIA protein levels. Black bars indicate no treatment control, red bars represent 0.5 hours of IL6 treatment, and blue bars represent 2 hours of IL6 treatment. ( C ) Knockdown of AMPK by shRNA reduces IL6-induced PFKL-upregulated protein expression in PLC5 cells, leading to decreases in pAMPK, PFKL, RPIA, pERK, and pSMAD5 levels, while it has no effect on STAT3 phosphorylation. The right panel illustrates the quantification of western blot results for AMPK, pAMPK, PFKL, RPIA, STAT3, pSTAT3, ERK, pERK, mTOR, p-mTOR, SMAD5, and pSMAD5. Black bars represent no treatment control, red bars indicate IL6 treatment, blue bars represent shAMPK, and the green bar denotes shAMPK+IL6. ( D ) Knockdown of AMPK decreases cell viability in PLC5 cells. Quantification of cell viability at 24, 48, and 72 hours is shown. Red bars represent shAMPK, while black bars denote shLuc control. ( E ) AMPK knockdown reduces IL6-stimulated cell viability, normalized to the control without IL6 treatment. Black bars represent the control without treatment, and red bars represent IL6 treatment. ( F ) Inhibition of AMPK with dorsomorphin decreases cell viability in three hepatoma cell lines. Cell death rates were quantified, and the IC50 for dorsomorphin in PLC5, Hep3B, and HepG2 cells is displayed in the upper left. ( G ) AMPK knockdown decreases migration ability with or without IL6 treatment. Quantification of migration without IL6 or with IL6 treatment is shown. Black bars represent sh-Luc control, and red bars denote sh-AMPK. ( H ) Suppression of AMPK with dorsomorphin reduces migration ability. Black bars represent the control without treatment, and red bars indicate dorsomorphin treatment. ( I ) Dorsomorphin significantly reduces PFKL protein levels. The left panel presents quantification of western blot results for AMPK, pAMPK, PFKL, SMAD5, pSMAD5, ACC, and pACC. Black bars represent the control without treatment, light red bars denote 1 µg/mL, and red bars indicate 6 µg/mL dorsomorphin treatment. ( J ) AMPK knockdown does not affect PFKL mRNA expression. Quantification of qPCR results for PFKL mRNA is shown. The red bar indicates dorsomorphin treatment, while the black bar denotes the control without treatment. ( K ) AMPK knockdown increases proteasome activity in PLC5 cells, suggesting that AMPK stabilizes PFKL by inhibiting proteasome activity. Quantification of proteasome activity is shown, with the red bar indicating shAMPK and the black bar representing shLuc control. Statistical analyses were performed using one-way ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: The specific antibodies against PFKL (Cell Signaling Technology Cat# 8175, RRID:AB_11178807), STAT3 (Cell Signaling Technology Cat# 9132, RRID:AB_331588), pSTAT3 (Cell Signaling Technology Cat# 9130, RRID:AB_330367), AMPK (Cell Signaling Technology Cat# 2603, RRID:AB_490795), pAMPK (Cell Signaling Technology Cat# 5759, RRID:AB_10949320), ERK (GeneTex Cat# GTX59618, RRID:AB_10726211), pERK (Abcam Cat# ab32538, RRID:AB_11156273), RPIA (Abcam Cat# ab67080, RRID:AB_1142656), PK (GeneTex Cat# GTX111536, RRID:AB_1951258), G6P (GeneTex Cat# GTX113203, RRID:AB_2037119), TPI (GeneTex Cat# GTX104618, RRID:AB_1241405), pRaf (BioVision Cat# 3504-100, RRID:AB_2060496), pMEK1/2 (Cell Signaling Technology Cat# 9121, RRID:AB_331648), pSMAD5 (Abcam Cat# ab92698, RRID:AB_10561456), SMAD5 (Abcam Cat# ab40771, RRID:AB_777981), pACC (Cell Signaling Technology Cat# 3661, AB_330337), ACC (Cell Signaling Technology Cat# 3662, RRID:AB_2219400), pmTOR(Cell Signaling Technology Cat#2974, RRID: AB_2262884), mTOR (Cell Signaling Technology Cat#2983, RRID: AB_ 2105622), α/β-Tubulin (Cell Signaling Technology Cat#2148, RRID: AB_2288042), GAPDH (GeneTex Cat# GTX100118, RRID:AB_1080976), β-actin (GeneTex Cat# GTX109639, RRID:AB_1949572) and ubiquitin (Cell Signaling Technology Cat# 3936, RRID:AB_331292) were purchased from Cell Signaling (Danvers, Massachusetts, USA), Abcam (Cambridge, Massachusetts, USA), and GeneTex (Irvine, CA, USA).

Techniques: Expressing, Control, Western Blot, Knockdown, shRNA, Phospho-proteomics, Inhibition, Migration, Activity Assay