pdk4 Search Results


gene exp pdk4 mm00443325 m1  (Thermo Fisher)
97
Thermo Fisher gene exp pdk4 mm00443325 m1
Analysis of mouse metabolic gene expression in 3T3-L1 adipocytes treated with NDP-MSH
Gene Exp Pdk4 Mm00443325 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gene exp pdk4 mm00443325 m1/product/Thermo Fisher
Average 97 stars, based on 1 article reviews
gene exp pdk4 mm00443325 m1 - by Bioz Stars, 2026-02
97/100 stars
  Buy from Supplier
anti pdk4 antibody  (Novus Biologicals)
93
Novus Biologicals anti pdk4 antibody
Expression of <t>PDK4</t> in siRNA-treated bladder cancer cell lines. ( A ) Fold change of PDK4 mRNA expression in T24 and J82 cells. ( B ) Protein expression of PDK4 in T24 and J82 cells. All data represent means ± SD of three independent experiments (* p < 0.05, ** p < 0.01 NC vs. T). M: mock, N: negative control, T: PDK4 siRNA treated.
Anti Pdk4 Antibody, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti pdk4 antibody/product/Novus Biologicals
Average 93 stars, based on 1 article reviews
anti pdk4 antibody - by Bioz Stars, 2026-02
93/100 stars
  Buy from Supplier
anti pdk4  (Santa Cruz Biotechnology)
93
Santa Cruz Biotechnology anti pdk4
Expression of <t>PDK4</t> in siRNA-treated bladder cancer cell lines. ( A ) Fold change of PDK4 mRNA expression in T24 and J82 cells. ( B ) Protein expression of PDK4 in T24 and J82 cells. All data represent means ± SD of three independent experiments (* p < 0.05, ** p < 0.01 NC vs. T). M: mock, N: negative control, T: PDK4 siRNA treated.
Anti Pdk4, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti pdk4/product/Santa Cruz Biotechnology
Average 93 stars, based on 1 article reviews
anti pdk4 - by Bioz Stars, 2026-02
93/100 stars
  Buy from Supplier
antibodies for pdk4  (Proteintech)
95
Proteintech antibodies for pdk4
FIGURE 4 | Protein–protein interactions of cardiac mitochondrial proteome. (A–C) Canonical pathways regulated in Y vs. O, Spm vs. O, and Spd vs. O groups. (D–F) STRING PPI analysis of differentially expressed proteins in Y vs. O, Spm vs. O, and Spd vs. O. (G) Construction of interaction network of <t>PDK4</t> by the String database.
Antibodies For Pdk4, 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/result/antibodies for pdk4/product/Proteintech
Average 95 stars, based on 1 article reviews
antibodies for pdk4 - by Bioz Stars, 2026-02
95/100 stars
  Buy from Supplier
pdk4  (OriGene)
90
OriGene pdk4
The RRC is regulated by pyruvate dehydrogenase kinases. ( a – c ) Neonatal rat cardiac myocytes were incubated for 24 h in complete growth medium under normoxic (atmospheric O 2 ) ( a ) or hypoxic (<1% O 2 ) ( b and c ) conditions. After this period, RNA was either immediately extracted and subjected to qPCR for the indicated genes ( b ), or the medium was replaced with one that is glucose and fatty acid free, or one containing 17.5 mM glucose, 100 μ M palmitate-BSA, or glucose+palmitate-BSA, as indicated, in normoxic conditions, for another 24 h ( a and c ). The results were averaged plotted as relative values to those from cells incubated in base medium in normoxic conditions adjusted to 1 ( a and c ) or control normoxia adjusted to 1 ( b ), n =3. Error bars represent standard error of the mean (S.E.M.), * P <0.05 versus control. ( d ) Neonatal cardiac myocyte was incubated for 24 h in complete growth medium and then infected with a control or adenoviruses (Ad) harboring PDK1, <t>PDK4</t> or Hif-1 α , for 24 h. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose plus 100 μ M palmitate for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed twice. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for control at the time point indicated; # P <0.05 max OCR for control versus max OCR of Pdk1 or Hif-1 α treated. ( e ) Neonatal cardiac myocytes were cultured for 24 h in complete growth medium containing vehicle or 1 μ M etomoxir, a Cpt1 inhibitor. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose plus 100 μ M palmitate containing vehicle or 1 μ M etomoxir, for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed twice. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for control at the time point indicated; # P <0.05 max OCR for control versus max OCR for etomoxir treated, at the time point indicated ( f and g ). Neonatal rat cardiac myocytes were cultured in complete growth medium containing vehicle or 1 mM DCA and either remained in normoxic conditions (atmospheric O 2 ) ( f ) or were exposed to hypoxia (<1% O 2 ) ( g ), for 24 h. At the end of this period, the medium was changed to base medium containing 17.5 mM glucose or 17.5 mM glucose plus 100 μ M palmitate-BSA containing vehicle or DCA, as indicated, for an additional 24 h. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose or 17.5 mM glucose plus 100 μ M palmitate-BSA containing vehicle or 1 mM DCA, for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed three times. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for DCA-treated cells, at the time point indicated; # P <0.05 max OCR for DCA-treated versus max OCR for untreated, at the time point indicated
Pdk4, supplied by OriGene, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pdk4/product/OriGene
Average 90 stars, based on 1 article reviews
pdk4 - by Bioz Stars, 2026-02
90/100 stars
  Buy from Supplier
gene exp pdk4 rn00585577 m1  (Thermo Fisher)
86
Thermo Fisher gene exp pdk4 rn00585577 m1
The RRC is regulated by pyruvate dehydrogenase kinases. ( a – c ) Neonatal rat cardiac myocytes were incubated for 24 h in complete growth medium under normoxic (atmospheric O 2 ) ( a ) or hypoxic (<1% O 2 ) ( b and c ) conditions. After this period, RNA was either immediately extracted and subjected to qPCR for the indicated genes ( b ), or the medium was replaced with one that is glucose and fatty acid free, or one containing 17.5 mM glucose, 100 μ M palmitate-BSA, or glucose+palmitate-BSA, as indicated, in normoxic conditions, for another 24 h ( a and c ). The results were averaged plotted as relative values to those from cells incubated in base medium in normoxic conditions adjusted to 1 ( a and c ) or control normoxia adjusted to 1 ( b ), n =3. Error bars represent standard error of the mean (S.E.M.), * P <0.05 versus control. ( d ) Neonatal cardiac myocyte was incubated for 24 h in complete growth medium and then infected with a control or adenoviruses (Ad) harboring PDK1, <t>PDK4</t> or Hif-1 α , for 24 h. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose plus 100 μ M palmitate for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed twice. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for control at the time point indicated; # P <0.05 max OCR for control versus max OCR of Pdk1 or Hif-1 α treated. ( e ) Neonatal cardiac myocytes were cultured for 24 h in complete growth medium containing vehicle or 1 μ M etomoxir, a Cpt1 inhibitor. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose plus 100 μ M palmitate containing vehicle or 1 μ M etomoxir, for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed twice. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for control at the time point indicated; # P <0.05 max OCR for control versus max OCR for etomoxir treated, at the time point indicated ( f and g ). Neonatal rat cardiac myocytes were cultured in complete growth medium containing vehicle or 1 mM DCA and either remained in normoxic conditions (atmospheric O 2 ) ( f ) or were exposed to hypoxia (<1% O 2 ) ( g ), for 24 h. At the end of this period, the medium was changed to base medium containing 17.5 mM glucose or 17.5 mM glucose plus 100 μ M palmitate-BSA containing vehicle or DCA, as indicated, for an additional 24 h. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose or 17.5 mM glucose plus 100 μ M palmitate-BSA containing vehicle or 1 mM DCA, for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed three times. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for DCA-treated cells, at the time point indicated; # P <0.05 max OCR for DCA-treated versus max OCR for untreated, at the time point indicated
Gene Exp Pdk4 Rn00585577 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gene exp pdk4 rn00585577 m1/product/Thermo Fisher
Average 86 stars, based on 1 article reviews
gene exp pdk4 rn00585577 m1 - by Bioz Stars, 2026-02
86/100 stars
  Buy from Supplier
gene exp pdk4 mm01166879 m1  (Thermo Fisher)
97
Thermo Fisher gene exp pdk4 mm01166879 m1
The RRC is regulated by pyruvate dehydrogenase kinases. ( a – c ) Neonatal rat cardiac myocytes were incubated for 24 h in complete growth medium under normoxic (atmospheric O 2 ) ( a ) or hypoxic (<1% O 2 ) ( b and c ) conditions. After this period, RNA was either immediately extracted and subjected to qPCR for the indicated genes ( b ), or the medium was replaced with one that is glucose and fatty acid free, or one containing 17.5 mM glucose, 100 μ M palmitate-BSA, or glucose+palmitate-BSA, as indicated, in normoxic conditions, for another 24 h ( a and c ). The results were averaged plotted as relative values to those from cells incubated in base medium in normoxic conditions adjusted to 1 ( a and c ) or control normoxia adjusted to 1 ( b ), n =3. Error bars represent standard error of the mean (S.E.M.), * P <0.05 versus control. ( d ) Neonatal cardiac myocyte was incubated for 24 h in complete growth medium and then infected with a control or adenoviruses (Ad) harboring PDK1, <t>PDK4</t> or Hif-1 α , for 24 h. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose plus 100 μ M palmitate for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed twice. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for control at the time point indicated; # P <0.05 max OCR for control versus max OCR of Pdk1 or Hif-1 α treated. ( e ) Neonatal cardiac myocytes were cultured for 24 h in complete growth medium containing vehicle or 1 μ M etomoxir, a Cpt1 inhibitor. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose plus 100 μ M palmitate containing vehicle or 1 μ M etomoxir, for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed twice. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for control at the time point indicated; # P <0.05 max OCR for control versus max OCR for etomoxir treated, at the time point indicated ( f and g ). Neonatal rat cardiac myocytes were cultured in complete growth medium containing vehicle or 1 mM DCA and either remained in normoxic conditions (atmospheric O 2 ) ( f ) or were exposed to hypoxia (<1% O 2 ) ( g ), for 24 h. At the end of this period, the medium was changed to base medium containing 17.5 mM glucose or 17.5 mM glucose plus 100 μ M palmitate-BSA containing vehicle or DCA, as indicated, for an additional 24 h. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose or 17.5 mM glucose plus 100 μ M palmitate-BSA containing vehicle or 1 mM DCA, for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed three times. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for DCA-treated cells, at the time point indicated; # P <0.05 max OCR for DCA-treated versus max OCR for untreated, at the time point indicated
Gene Exp Pdk4 Mm01166879 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gene exp pdk4 mm01166879 m1/product/Thermo Fisher
Average 97 stars, based on 1 article reviews
gene exp pdk4 mm01166879 m1 - by Bioz Stars, 2026-02
97/100 stars
  Buy from Supplier
description sequence pyy10  (Addgene inc)
93
Addgene inc description sequence pyy10
The RRC is regulated by pyruvate dehydrogenase kinases. ( a – c ) Neonatal rat cardiac myocytes were incubated for 24 h in complete growth medium under normoxic (atmospheric O 2 ) ( a ) or hypoxic (<1% O 2 ) ( b and c ) conditions. After this period, RNA was either immediately extracted and subjected to qPCR for the indicated genes ( b ), or the medium was replaced with one that is glucose and fatty acid free, or one containing 17.5 mM glucose, 100 μ M palmitate-BSA, or glucose+palmitate-BSA, as indicated, in normoxic conditions, for another 24 h ( a and c ). The results were averaged plotted as relative values to those from cells incubated in base medium in normoxic conditions adjusted to 1 ( a and c ) or control normoxia adjusted to 1 ( b ), n =3. Error bars represent standard error of the mean (S.E.M.), * P <0.05 versus control. ( d ) Neonatal cardiac myocyte was incubated for 24 h in complete growth medium and then infected with a control or adenoviruses (Ad) harboring PDK1, <t>PDK4</t> or Hif-1 α , for 24 h. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose plus 100 μ M palmitate for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed twice. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for control at the time point indicated; # P <0.05 max OCR for control versus max OCR of Pdk1 or Hif-1 α treated. ( e ) Neonatal cardiac myocytes were cultured for 24 h in complete growth medium containing vehicle or 1 μ M etomoxir, a Cpt1 inhibitor. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose plus 100 μ M palmitate containing vehicle or 1 μ M etomoxir, for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed twice. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for control at the time point indicated; # P <0.05 max OCR for control versus max OCR for etomoxir treated, at the time point indicated ( f and g ). Neonatal rat cardiac myocytes were cultured in complete growth medium containing vehicle or 1 mM DCA and either remained in normoxic conditions (atmospheric O 2 ) ( f ) or were exposed to hypoxia (<1% O 2 ) ( g ), for 24 h. At the end of this period, the medium was changed to base medium containing 17.5 mM glucose or 17.5 mM glucose plus 100 μ M palmitate-BSA containing vehicle or DCA, as indicated, for an additional 24 h. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose or 17.5 mM glucose plus 100 μ M palmitate-BSA containing vehicle or 1 mM DCA, for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed three times. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for DCA-treated cells, at the time point indicated; # P <0.05 max OCR for DCA-treated versus max OCR for untreated, at the time point indicated
Description Sequence Pyy10, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/description sequence pyy10/product/Addgene inc
Average 93 stars, based on 1 article reviews
description sequence pyy10 - by Bioz Stars, 2026-02
93/100 stars
  Buy from Supplier
pdk4  (Atlas Antibodies)
93
Atlas Antibodies pdk4
Figure 1. Expression of PDK2 and <t>PDK4</t> in hindpaw tissues after CFA injection. A, The expression of Pdk2 and Pdk4 mRNAs in hindpaw tissues at different time points after CFA injection was assessed by real-time RT-PCR. Pdk2 and Pdk4 mRNA levels in CFA-injectedhindpawtissuesweresignificantlyupregulatedafterinjection,peakingat3dandthensubsiding.ResultsformRNA expression are displayed as the fold increase of gene expression normalized to GAPDH. B, Protein levels of PDK2 and PDK4 in hindpaw tissues 3 d after CFA injection were assessed by Western blot analysis. Quantifications of the band intensities are pre- sentedintheadjacentgraphs.C,D,ImmunofluorescenceanalysisdetectedastrongexpressionofPDK2andPDK4inthehindpaw tissues of CFA-injected mice at 3 d after injection, but not in the vehicle-treated control animals. *p 0.05 versus the vehicle- treated control animals (Student’s t test). n 3. Data are mean SEM. Scale bars, 200 m. Images show the representative results of at least three independent experiments.
Pdk4, supplied by Atlas Antibodies, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pdk4/product/Atlas Antibodies
Average 93 stars, based on 1 article reviews
pdk4 - by Bioz Stars, 2026-02
93/100 stars
  Buy from Supplier
pdk4 inhibitor 8c  (MedChemExpress)
93
MedChemExpress pdk4 inhibitor 8c
Figure 1. Expression of PDK2 and <t>PDK4</t> in hindpaw tissues after CFA injection. A, The expression of Pdk2 and Pdk4 mRNAs in hindpaw tissues at different time points after CFA injection was assessed by real-time RT-PCR. Pdk2 and Pdk4 mRNA levels in CFA-injectedhindpawtissuesweresignificantlyupregulatedafterinjection,peakingat3dandthensubsiding.ResultsformRNA expression are displayed as the fold increase of gene expression normalized to GAPDH. B, Protein levels of PDK2 and PDK4 in hindpaw tissues 3 d after CFA injection were assessed by Western blot analysis. Quantifications of the band intensities are pre- sentedintheadjacentgraphs.C,D,ImmunofluorescenceanalysisdetectedastrongexpressionofPDK2andPDK4inthehindpaw tissues of CFA-injected mice at 3 d after injection, but not in the vehicle-treated control animals. *p 0.05 versus the vehicle- treated control animals (Student’s t test). n 3. Data are mean SEM. Scale bars, 200 m. Images show the representative results of at least three independent experiments.
Pdk4 Inhibitor 8c, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pdk4 inhibitor 8c/product/MedChemExpress
Average 93 stars, based on 1 article reviews
pdk4 inhibitor 8c - by Bioz Stars, 2026-02
93/100 stars
  Buy from Supplier
pdk4  (Novus Biologicals)
92
Novus Biologicals pdk4
Quantitative PCR primer sequences
Pdk4, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pdk4/product/Novus Biologicals
Average 92 stars, based on 1 article reviews
pdk4 - by Bioz Stars, 2026-02
92/100 stars
  Buy from Supplier

Image Search Results


Analysis of mouse metabolic gene expression in 3T3-L1 adipocytes treated with NDP-MSH

Journal: Molecular Endocrinology

Article Title: Nr4a1 siRNA Expression Attenuates α-MSH Regulated Gene Expression in 3T3-L1 Adipocytes

doi: 10.1210/me.2010-0231

Figure Lengend Snippet: Analysis of mouse metabolic gene expression in 3T3-L1 adipocytes treated with NDP-MSH

Article Snippet: Pdk4-Mm00443325_m1 , 2.01954 , − 11.3726813856132 , 0.00008 , 0.00171 , 2.59628 , 0.41435 , Valid.

Techniques: Gene Expression

Analysis of mouse metabolic gene expression in 3T3-L1 adipocytes treated with 8-Br cAMP

Journal: Molecular Endocrinology

Article Title: Nr4a1 siRNA Expression Attenuates α-MSH Regulated Gene Expression in 3T3-L1 Adipocytes

doi: 10.1210/me.2010-0231

Figure Lengend Snippet: Analysis of mouse metabolic gene expression in 3T3-L1 adipocytes treated with 8-Br cAMP

Article Snippet: Pdk4-Mm00443325_m1 , 2.01954 , − 11.3726813856132 , 0.00008 , 0.00171 , 2.59628 , 0.41435 , Valid.

Techniques: Gene Expression

The NDP-MSH induction of the Nr4a subgroup is concomitant with significant differential expression genes involved in metabolism. Total RNA from 3T3-L1 adipocytes treated with 10 nM NDP-MSH from 15 min to 8 h were analyzed by qPCR for the expression of Il6 (A), Cox2/Ptgs2 (B), Pck1 (C), and Pdk4 (D). Results were normalized against 36b4 at each time point and expressed as mean ± sem (n = 3). E, IL-6 secretion to medium (pictograms per milliliter). Medium from 3T3-L1 adipocytes treated for 2 h with 10 nm NDP-MSH and/or vehicle was collected and measured. Results are mean ± sem of three independent experiments performed in quadruplet. Statistical significance was calculated using the Student's unpaired t test, where *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Journal: Molecular Endocrinology

Article Title: Nr4a1 siRNA Expression Attenuates α-MSH Regulated Gene Expression in 3T3-L1 Adipocytes

doi: 10.1210/me.2010-0231

Figure Lengend Snippet: The NDP-MSH induction of the Nr4a subgroup is concomitant with significant differential expression genes involved in metabolism. Total RNA from 3T3-L1 adipocytes treated with 10 nM NDP-MSH from 15 min to 8 h were analyzed by qPCR for the expression of Il6 (A), Cox2/Ptgs2 (B), Pck1 (C), and Pdk4 (D). Results were normalized against 36b4 at each time point and expressed as mean ± sem (n = 3). E, IL-6 secretion to medium (pictograms per milliliter). Medium from 3T3-L1 adipocytes treated for 2 h with 10 nm NDP-MSH and/or vehicle was collected and measured. Results are mean ± sem of three independent experiments performed in quadruplet. Statistical significance was calculated using the Student's unpaired t test, where *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: Pdk4-Mm00443325_m1 , 2.01954 , − 11.3726813856132 , 0.00008 , 0.00171 , 2.59628 , 0.41435 , Valid.

Techniques: Quantitative Proteomics, Expressing

Nur77 siRNA stable expression significantly attenuates the NDP-MSH-mediated induction of Nr4a subgroup and metabolic genes. 3T3-L1 Nur77 siRNA (siNur77) and negative siRNA (siNeg) stable cells were differentiated and treated with 10 nm NDP-MSH or vehicle for 2 h. qPCR was used to assay the expression of Nur77/Nr4a1 (A), Nurr1/Nr4a2 (B), Nor-1/Nr4a3 (C), Il6 (D), Cox2/Ptgs2 (E), Pck1 (F), and Pdk4 (G). Results were normalized against 36b4 and expressed as fold changes (n = 3). Statistical significance was calculated using the Student's unpaired t test, where *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Journal: Molecular Endocrinology

Article Title: Nr4a1 siRNA Expression Attenuates α-MSH Regulated Gene Expression in 3T3-L1 Adipocytes

doi: 10.1210/me.2010-0231

Figure Lengend Snippet: Nur77 siRNA stable expression significantly attenuates the NDP-MSH-mediated induction of Nr4a subgroup and metabolic genes. 3T3-L1 Nur77 siRNA (siNur77) and negative siRNA (siNeg) stable cells were differentiated and treated with 10 nm NDP-MSH or vehicle for 2 h. qPCR was used to assay the expression of Nur77/Nr4a1 (A), Nurr1/Nr4a2 (B), Nor-1/Nr4a3 (C), Il6 (D), Cox2/Ptgs2 (E), Pck1 (F), and Pdk4 (G). Results were normalized against 36b4 and expressed as fold changes (n = 3). Statistical significance was calculated using the Student's unpaired t test, where *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: Pdk4-Mm00443325_m1 , 2.01954 , − 11.3726813856132 , 0.00008 , 0.00171 , 2.59628 , 0.41435 , Valid.

Techniques: Expressing

NDP-MSH induces Nur77, Il6, Cox2, Pck1, and Pdk4 gene expressions in mouse adipose tissue. Mice were ip injected with vehicle (saline) or 1 mg/kg of NDP-MSH and inguinal adipose tissue collected after 4 h. qPCR was used to assay the mRNA levels, and results were normalized against 36b4 and expressed as fold changes (n = 7–8). Statistical significance was calculated using the Mann-Whitney U test, where *, P < 0.05 and ***, P < 0.001.

Journal: Molecular Endocrinology

Article Title: Nr4a1 siRNA Expression Attenuates α-MSH Regulated Gene Expression in 3T3-L1 Adipocytes

doi: 10.1210/me.2010-0231

Figure Lengend Snippet: NDP-MSH induces Nur77, Il6, Cox2, Pck1, and Pdk4 gene expressions in mouse adipose tissue. Mice were ip injected with vehicle (saline) or 1 mg/kg of NDP-MSH and inguinal adipose tissue collected after 4 h. qPCR was used to assay the mRNA levels, and results were normalized against 36b4 and expressed as fold changes (n = 7–8). Statistical significance was calculated using the Mann-Whitney U test, where *, P < 0.05 and ***, P < 0.001.

Article Snippet: Pdk4-Mm00443325_m1 , 2.01954 , − 11.3726813856132 , 0.00008 , 0.00171 , 2.59628 , 0.41435 , Valid.

Techniques: Injection, Saline, MANN-WHITNEY

Expression of PDK4 in siRNA-treated bladder cancer cell lines. ( A ) Fold change of PDK4 mRNA expression in T24 and J82 cells. ( B ) Protein expression of PDK4 in T24 and J82 cells. All data represent means ± SD of three independent experiments (* p < 0.05, ** p < 0.01 NC vs. T). M: mock, N: negative control, T: PDK4 siRNA treated.

Journal: International Journal of Molecular Sciences

Article Title: Anti-Metastatic Effect of Pyruvate Dehydrogenase Kinase 4 Inhibition in Bladder Cancer via the ERK, SRC, and JNK Pathways

doi: 10.3390/ijms232113240

Figure Lengend Snippet: Expression of PDK4 in siRNA-treated bladder cancer cell lines. ( A ) Fold change of PDK4 mRNA expression in T24 and J82 cells. ( B ) Protein expression of PDK4 in T24 and J82 cells. All data represent means ± SD of three independent experiments (* p < 0.05, ** p < 0.01 NC vs. T). M: mock, N: negative control, T: PDK4 siRNA treated.

Article Snippet: Anti-PDK4 antibody was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Expressing, Negative Control

Migration and invasion assay of PDK4 knockdown bladder cancer cells. ( A ) Migration assay. ( B ) Invasion assay. All data represent means ± SD of three independent experiments (* p < 0.05, ** p < 0.01 NC vs. T). M: mock, N: negative control, T: PDK4 siRNA treated.

Journal: International Journal of Molecular Sciences

Article Title: Anti-Metastatic Effect of Pyruvate Dehydrogenase Kinase 4 Inhibition in Bladder Cancer via the ERK, SRC, and JNK Pathways

doi: 10.3390/ijms232113240

Figure Lengend Snippet: Migration and invasion assay of PDK4 knockdown bladder cancer cells. ( A ) Migration assay. ( B ) Invasion assay. All data represent means ± SD of three independent experiments (* p < 0.05, ** p < 0.01 NC vs. T). M: mock, N: negative control, T: PDK4 siRNA treated.

Article Snippet: Anti-PDK4 antibody was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Migration, Invasion Assay, Knockdown, Negative Control

PDK4-related protein expression in PDK4 knockdown bladder cancer cells. β-actin is used as an internal control. M: mock, N: negative control, T: PDK4 siRNA treated. All data represent means ± SD of three independent experiments (* p < 0.05, ** p < 0.01 NC vs. T). M: mock, N: negative control, T: PDK4 siRNA treated.

Journal: International Journal of Molecular Sciences

Article Title: Anti-Metastatic Effect of Pyruvate Dehydrogenase Kinase 4 Inhibition in Bladder Cancer via the ERK, SRC, and JNK Pathways

doi: 10.3390/ijms232113240

Figure Lengend Snippet: PDK4-related protein expression in PDK4 knockdown bladder cancer cells. β-actin is used as an internal control. M: mock, N: negative control, T: PDK4 siRNA treated. All data represent means ± SD of three independent experiments (* p < 0.05, ** p < 0.01 NC vs. T). M: mock, N: negative control, T: PDK4 siRNA treated.

Article Snippet: Anti-PDK4 antibody was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Expressing, Knockdown, Control, Negative Control

Xenograft model of PDK4 knockdown J82 cells. ( A ) Tumor growth. (* p < 0.05, ** p < 0.01 Day 0 vs. Each day). Ctrl: wild type, KD: knockdown. ( B ) Gross appearance. ( C ) Representative image of immunohisto-chemistry. (* p < 0.05, ** p < 0.01 Ctrl vs. KD).

Journal: International Journal of Molecular Sciences

Article Title: Anti-Metastatic Effect of Pyruvate Dehydrogenase Kinase 4 Inhibition in Bladder Cancer via the ERK, SRC, and JNK Pathways

doi: 10.3390/ijms232113240

Figure Lengend Snippet: Xenograft model of PDK4 knockdown J82 cells. ( A ) Tumor growth. (* p < 0.05, ** p < 0.01 Day 0 vs. Each day). Ctrl: wild type, KD: knockdown. ( B ) Gross appearance. ( C ) Representative image of immunohisto-chemistry. (* p < 0.05, ** p < 0.01 Ctrl vs. KD).

Article Snippet: Anti-PDK4 antibody was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Knockdown, Immunohistochemistry

PDK4 expression in human bladder cancer specimen. ( A ) Relative PDK4 mRNA expression in normal and bladder cancer specimen. ( B ) Representative image of immunohisto-chemistry of PDK4 protein. All data represent means ± SD of three independent experiments (** p < 0.01 normal vs. T stages).

Journal: International Journal of Molecular Sciences

Article Title: Anti-Metastatic Effect of Pyruvate Dehydrogenase Kinase 4 Inhibition in Bladder Cancer via the ERK, SRC, and JNK Pathways

doi: 10.3390/ijms232113240

Figure Lengend Snippet: PDK4 expression in human bladder cancer specimen. ( A ) Relative PDK4 mRNA expression in normal and bladder cancer specimen. ( B ) Representative image of immunohisto-chemistry of PDK4 protein. All data represent means ± SD of three independent experiments (** p < 0.01 normal vs. T stages).

Article Snippet: Anti-PDK4 antibody was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Expressing, Immunohistochemistry

MS-based quantitative proteomic profiling of phosphorylation in bladder cancer cell lines. ( A ) Experimental design of proteomic analysis. J82 and J82 KD cells were lysed, subjected to in-solution digestion and 18 O labeling, and then mixed at equal protein amounts. Phosphorylated peptides were enriched by TiO 2 . Eluted peptides were analysis using LC-MS/MS. Mass spectrum data were searched in the MaxQuant (version 1.5) database. ( B ) Venn diagrams showing overlap between J82 knock down and J82 control. ( C ) Volcano plot for 209 differentially phosphorylated protein (DRPs) between PDK4 knockdown and controls.

Journal: International Journal of Molecular Sciences

Article Title: Anti-Metastatic Effect of Pyruvate Dehydrogenase Kinase 4 Inhibition in Bladder Cancer via the ERK, SRC, and JNK Pathways

doi: 10.3390/ijms232113240

Figure Lengend Snippet: MS-based quantitative proteomic profiling of phosphorylation in bladder cancer cell lines. ( A ) Experimental design of proteomic analysis. J82 and J82 KD cells were lysed, subjected to in-solution digestion and 18 O labeling, and then mixed at equal protein amounts. Phosphorylated peptides were enriched by TiO 2 . Eluted peptides were analysis using LC-MS/MS. Mass spectrum data were searched in the MaxQuant (version 1.5) database. ( B ) Venn diagrams showing overlap between J82 knock down and J82 control. ( C ) Volcano plot for 209 differentially phosphorylated protein (DRPs) between PDK4 knockdown and controls.

Article Snippet: Anti-PDK4 antibody was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Phospho-proteomics, Labeling, Liquid Chromatography with Mass Spectroscopy, Knockdown, Control

FIGURE 4 | Protein–protein interactions of cardiac mitochondrial proteome. (A–C) Canonical pathways regulated in Y vs. O, Spm vs. O, and Spd vs. O groups. (D–F) STRING PPI analysis of differentially expressed proteins in Y vs. O, Spm vs. O, and Spd vs. O. (G) Construction of interaction network of PDK4 by the String database.

Journal: Frontiers in cell and developmental biology

Article Title: Dynamic Mitochondrial Proteome Under Polyamines Treatment in Cardiac Aging.

doi: 10.3389/fcell.2022.840389

Figure Lengend Snippet: FIGURE 4 | Protein–protein interactions of cardiac mitochondrial proteome. (A–C) Canonical pathways regulated in Y vs. O, Spm vs. O, and Spd vs. O groups. (D–F) STRING PPI analysis of differentially expressed proteins in Y vs. O, Spm vs. O, and Spd vs. O. (G) Construction of interaction network of PDK4 by the String database.

Article Snippet: The following antibodies were used: Antibodies for PDK4 (Cat# 12949-1-AP), HADHA (Cat# 10758-1-AP), Annexin 6 (Cat# 12542-1-AP), NNT (Cat# 13442-2-AP), and COX-IV (Cat# 66110-1-Ig) were obtained from Proteintech (Wuhan, Hubei, China).

Techniques: Protein-Protein interactions

FIGURE 5 | Validation of differentially expressed proteins. (A–D) WB analysis of PDK4, NNT, Annexin 6, and HADHA protein level in myocardial tissue of Y, O, Spm, or Spd-treated rats (top); histogram shows the expression levels of the four proteins as determined by densitometric analysis (bottom), mitochondrial markers protein (COX IV) is used as the internal loading control (n = 6). Mean ± SEM; p-values correspond to one-way ANOVA; *p < 0.05 vs. Y; §p < 0.05, and #p < 0.05 vs. O.

Journal: Frontiers in cell and developmental biology

Article Title: Dynamic Mitochondrial Proteome Under Polyamines Treatment in Cardiac Aging.

doi: 10.3389/fcell.2022.840389

Figure Lengend Snippet: FIGURE 5 | Validation of differentially expressed proteins. (A–D) WB analysis of PDK4, NNT, Annexin 6, and HADHA protein level in myocardial tissue of Y, O, Spm, or Spd-treated rats (top); histogram shows the expression levels of the four proteins as determined by densitometric analysis (bottom), mitochondrial markers protein (COX IV) is used as the internal loading control (n = 6). Mean ± SEM; p-values correspond to one-way ANOVA; *p < 0.05 vs. Y; §p < 0.05, and #p < 0.05 vs. O.

Article Snippet: The following antibodies were used: Antibodies for PDK4 (Cat# 12949-1-AP), HADHA (Cat# 10758-1-AP), Annexin 6 (Cat# 12542-1-AP), NNT (Cat# 13442-2-AP), and COX-IV (Cat# 66110-1-Ig) were obtained from Proteintech (Wuhan, Hubei, China).

Techniques: Biomarker Discovery, Expressing, Control

FIGURE 6 | Polyamines inhibited cardiac aging through downregulating PDK4. (A) Representative images showing the morphological changes and senescence- associated- β-galactosidase (SA-β-gal) activity in cardiomyocytes treated with H2O2, H2O2-Spm, H2O2-Spd, and H2O2-Spd-DFMO, scale bar = 20 μm. (B) Quantitative analysis of SA-β-gal-positive cells percentage (n = 4). (C) The expression of p21 and PDK4 protein evaluated by WB analysis. (D) Quantitative analysis of p21 and PDK4 level normalized to GAPDH (n = 4–6). (E) Representative images of JC-1staining in cardiomyocytes treated with H2O2, H2O2-Spm, H2O2-Spd, and H2O2-DAC (an inhibitor of PDK4), respectively. Red fluorescence represents JC-1 aggregates formed in normal cells with high Δψm, whereas green fluorescence represents JC-1 (Continued)

Journal: Frontiers in cell and developmental biology

Article Title: Dynamic Mitochondrial Proteome Under Polyamines Treatment in Cardiac Aging.

doi: 10.3389/fcell.2022.840389

Figure Lengend Snippet: FIGURE 6 | Polyamines inhibited cardiac aging through downregulating PDK4. (A) Representative images showing the morphological changes and senescence- associated- β-galactosidase (SA-β-gal) activity in cardiomyocytes treated with H2O2, H2O2-Spm, H2O2-Spd, and H2O2-Spd-DFMO, scale bar = 20 μm. (B) Quantitative analysis of SA-β-gal-positive cells percentage (n = 4). (C) The expression of p21 and PDK4 protein evaluated by WB analysis. (D) Quantitative analysis of p21 and PDK4 level normalized to GAPDH (n = 4–6). (E) Representative images of JC-1staining in cardiomyocytes treated with H2O2, H2O2-Spm, H2O2-Spd, and H2O2-DAC (an inhibitor of PDK4), respectively. Red fluorescence represents JC-1 aggregates formed in normal cells with high Δψm, whereas green fluorescence represents JC-1 (Continued)

Article Snippet: The following antibodies were used: Antibodies for PDK4 (Cat# 12949-1-AP), HADHA (Cat# 10758-1-AP), Annexin 6 (Cat# 12542-1-AP), NNT (Cat# 13442-2-AP), and COX-IV (Cat# 66110-1-Ig) were obtained from Proteintech (Wuhan, Hubei, China).

Techniques: Activity Assay, Expressing

The RRC is regulated by pyruvate dehydrogenase kinases. ( a – c ) Neonatal rat cardiac myocytes were incubated for 24 h in complete growth medium under normoxic (atmospheric O 2 ) ( a ) or hypoxic (<1% O 2 ) ( b and c ) conditions. After this period, RNA was either immediately extracted and subjected to qPCR for the indicated genes ( b ), or the medium was replaced with one that is glucose and fatty acid free, or one containing 17.5 mM glucose, 100 μ M palmitate-BSA, or glucose+palmitate-BSA, as indicated, in normoxic conditions, for another 24 h ( a and c ). The results were averaged plotted as relative values to those from cells incubated in base medium in normoxic conditions adjusted to 1 ( a and c ) or control normoxia adjusted to 1 ( b ), n =3. Error bars represent standard error of the mean (S.E.M.), * P <0.05 versus control. ( d ) Neonatal cardiac myocyte was incubated for 24 h in complete growth medium and then infected with a control or adenoviruses (Ad) harboring PDK1, PDK4 or Hif-1 α , for 24 h. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose plus 100 μ M palmitate for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed twice. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for control at the time point indicated; # P <0.05 max OCR for control versus max OCR of Pdk1 or Hif-1 α treated. ( e ) Neonatal cardiac myocytes were cultured for 24 h in complete growth medium containing vehicle or 1 μ M etomoxir, a Cpt1 inhibitor. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose plus 100 μ M palmitate containing vehicle or 1 μ M etomoxir, for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed twice. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for control at the time point indicated; # P <0.05 max OCR for control versus max OCR for etomoxir treated, at the time point indicated ( f and g ). Neonatal rat cardiac myocytes were cultured in complete growth medium containing vehicle or 1 mM DCA and either remained in normoxic conditions (atmospheric O 2 ) ( f ) or were exposed to hypoxia (<1% O 2 ) ( g ), for 24 h. At the end of this period, the medium was changed to base medium containing 17.5 mM glucose or 17.5 mM glucose plus 100 μ M palmitate-BSA containing vehicle or DCA, as indicated, for an additional 24 h. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose or 17.5 mM glucose plus 100 μ M palmitate-BSA containing vehicle or 1 mM DCA, for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed three times. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for DCA-treated cells, at the time point indicated; # P <0.05 max OCR for DCA-treated versus max OCR for untreated, at the time point indicated

Journal: Cell Death & Disease

Article Title: Mitochondrial complex II is a source of the reserve respiratory capacity that is regulated by metabolic sensors and promotes cell survival

doi: 10.1038/cddis.2015.202

Figure Lengend Snippet: The RRC is regulated by pyruvate dehydrogenase kinases. ( a – c ) Neonatal rat cardiac myocytes were incubated for 24 h in complete growth medium under normoxic (atmospheric O 2 ) ( a ) or hypoxic (<1% O 2 ) ( b and c ) conditions. After this period, RNA was either immediately extracted and subjected to qPCR for the indicated genes ( b ), or the medium was replaced with one that is glucose and fatty acid free, or one containing 17.5 mM glucose, 100 μ M palmitate-BSA, or glucose+palmitate-BSA, as indicated, in normoxic conditions, for another 24 h ( a and c ). The results were averaged plotted as relative values to those from cells incubated in base medium in normoxic conditions adjusted to 1 ( a and c ) or control normoxia adjusted to 1 ( b ), n =3. Error bars represent standard error of the mean (S.E.M.), * P <0.05 versus control. ( d ) Neonatal cardiac myocyte was incubated for 24 h in complete growth medium and then infected with a control or adenoviruses (Ad) harboring PDK1, PDK4 or Hif-1 α , for 24 h. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose plus 100 μ M palmitate for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed twice. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for control at the time point indicated; # P <0.05 max OCR for control versus max OCR of Pdk1 or Hif-1 α treated. ( e ) Neonatal cardiac myocytes were cultured for 24 h in complete growth medium containing vehicle or 1 μ M etomoxir, a Cpt1 inhibitor. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose plus 100 μ M palmitate containing vehicle or 1 μ M etomoxir, for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed twice. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for control at the time point indicated; # P <0.05 max OCR for control versus max OCR for etomoxir treated, at the time point indicated ( f and g ). Neonatal rat cardiac myocytes were cultured in complete growth medium containing vehicle or 1 mM DCA and either remained in normoxic conditions (atmospheric O 2 ) ( f ) or were exposed to hypoxia (<1% O 2 ) ( g ), for 24 h. At the end of this period, the medium was changed to base medium containing 17.5 mM glucose or 17.5 mM glucose plus 100 μ M palmitate-BSA containing vehicle or DCA, as indicated, for an additional 24 h. The medium was then replaced with serum-free XF medium containing 17.5 mM glucose or 17.5 mM glucose plus 100 μ M palmitate-BSA containing vehicle or 1 mM DCA, for 1 h. The mitochondrial stress test was then performed as described in Materials and Methods, n =4–6, each experiment was performed three times. Error bars represent S.E.M., * P <0.05 max OCR versus basal OCR for DCA-treated cells, at the time point indicated; # P <0.05 max OCR for DCA-treated versus max OCR for untreated, at the time point indicated

Article Snippet: The human cDNA clones for Pdk1 (SC321678/NM_002610), Pdk4 (SC118542/NM_002612), Sdhaf1 (NM_001042631) and Sirt3 (NM_012239) were purchased (Origene Technologies, Inc., Rockville, MD, USA) and cloned into recombinant adenovirus vectors, propagated, and titered as previously described by Graham and Prevec.

Techniques: Incubation, Infection, Cell Culture

Figure 1. Expression of PDK2 and PDK4 in hindpaw tissues after CFA injection. A, The expression of Pdk2 and Pdk4 mRNAs in hindpaw tissues at different time points after CFA injection was assessed by real-time RT-PCR. Pdk2 and Pdk4 mRNA levels in CFA-injectedhindpawtissuesweresignificantlyupregulatedafterinjection,peakingat3dandthensubsiding.ResultsformRNA expression are displayed as the fold increase of gene expression normalized to GAPDH. B, Protein levels of PDK2 and PDK4 in hindpaw tissues 3 d after CFA injection were assessed by Western blot analysis. Quantifications of the band intensities are pre- sentedintheadjacentgraphs.C,D,ImmunofluorescenceanalysisdetectedastrongexpressionofPDK2andPDK4inthehindpaw tissues of CFA-injected mice at 3 d after injection, but not in the vehicle-treated control animals. *p 0.05 versus the vehicle- treated control animals (Student’s t test). n 3. Data are mean SEM. Scale bars, 200 m. Images show the representative results of at least three independent experiments.

Journal: Journal of Neuroscience

Article Title: Metabolic Connection of Inflammatory Pain: Pivotal Role of a Pyruvate Dehydrogenase Kinase-Pyruvate Dehydrogenase-Lactic Acid Axis

doi: 10.1523/jneurosci.1910-15.2015

Figure Lengend Snippet: Figure 1. Expression of PDK2 and PDK4 in hindpaw tissues after CFA injection. A, The expression of Pdk2 and Pdk4 mRNAs in hindpaw tissues at different time points after CFA injection was assessed by real-time RT-PCR. Pdk2 and Pdk4 mRNA levels in CFA-injectedhindpawtissuesweresignificantlyupregulatedafterinjection,peakingat3dandthensubsiding.ResultsformRNA expression are displayed as the fold increase of gene expression normalized to GAPDH. B, Protein levels of PDK2 and PDK4 in hindpaw tissues 3 d after CFA injection were assessed by Western blot analysis. Quantifications of the band intensities are pre- sentedintheadjacentgraphs.C,D,ImmunofluorescenceanalysisdetectedastrongexpressionofPDK2andPDK4inthehindpaw tissues of CFA-injected mice at 3 d after injection, but not in the vehicle-treated control animals. *p 0.05 versus the vehicle- treated control animals (Student’s t test). n 3. Data are mean SEM. Scale bars, 200 m. Images show the representative results of at least three independent experiments.

Article Snippet: For immunofluorescence staining, sections were incubated with primary antibodies against PDK2 (rabbit, 1:200; Acris Antibodies), PDK4 (rabbit, 1:200; Atlas Antibodies), phospho-Ser 293-PDHE1 (pyruvate dehydrogenase E1 ) (rabbit, 1:200; Calbiochem), phospho-Ser 300-PDHE1 (pyruvate dehydrogenase E1 ) (rabbit, 1:200; Calbiochem), Iba-1 (rabbit, 1:1000; Wako; or goat, 1:200; Novus Biologicals [for costainings]), GFAP (mouse, 1:500; BD Biosciences), Ly6G (rat, 1:200; eBioscience) or iNOS (rabbit, 1:200; BD Biosciences), MYH1/2/3 (mouse, 1:200; Santa Cruz Biotechnology) and 3 tubulin (2G10) (mouse, 1:200; Santa Cruz Biotechnology), overnight at 4°C, and then incubated with FITC- or Cy3conjugated secondary antibodies (1:200; Jackson ImmunoResearch Laboratories).

Techniques: Expressing, Injection, Quantitative RT-PCR, Gene Expression, Western Blot, Control

Figure 5. Role of PDK2/4 in regulating the phenotypes of cultured macrophages. Peritoneal macrophage cultures preparedfromWTandPdk2/4DKOmiceweretreatedwithM1-phenotypeinducermixture[LPS(100ng/ml)plusIFN-(50 U/ml)] for 8 h. A, The mRNA levels of M1-related genes TNF-, IL-1, and IL-6 were assessed by real-time RT-PCR (left). Similarly, peritoneal macrophage cultures prepared from WT and Pdk2/4 DKO mice were treated with M2-phenotype inducer [IL-4 (10 ng/ml)] for 8 h, and the mRNA levels of M2-related genes Ym-1, Arg-1, and IL-10 were then assessed by real-time RT-PCR (right). B, The expression of IRF8 or IRF4 mRNAs in the cultured peritoneal macrophages (prepared from WT and Pdk2/4 DKO mice) following stimulation with LPS (100 ng/ml) plus IFN- (50 U/ml) or IL-4 (10 ng/ml) for 8 h was assessed by real-time RT-PCR. C, The expression of Pdk2 and Pdk4 mRNAs in the WT peritoneal macrophages following stimulation with LPS (100 ng/ml) plus IFN- (50 U/ml) for 8 h was assessed by real-time RT-PCR. Results for mRNA expression are displayed as the fold increase of gene expression normalized to GAPDH. *p 0.05 versus the control group. #p 0.05 between indicated groups (Student’s t test). n 3. Data are mean SEM.

Journal: Journal of Neuroscience

Article Title: Metabolic Connection of Inflammatory Pain: Pivotal Role of a Pyruvate Dehydrogenase Kinase-Pyruvate Dehydrogenase-Lactic Acid Axis

doi: 10.1523/jneurosci.1910-15.2015

Figure Lengend Snippet: Figure 5. Role of PDK2/4 in regulating the phenotypes of cultured macrophages. Peritoneal macrophage cultures preparedfromWTandPdk2/4DKOmiceweretreatedwithM1-phenotypeinducermixture[LPS(100ng/ml)plusIFN-(50 U/ml)] for 8 h. A, The mRNA levels of M1-related genes TNF-, IL-1, and IL-6 were assessed by real-time RT-PCR (left). Similarly, peritoneal macrophage cultures prepared from WT and Pdk2/4 DKO mice were treated with M2-phenotype inducer [IL-4 (10 ng/ml)] for 8 h, and the mRNA levels of M2-related genes Ym-1, Arg-1, and IL-10 were then assessed by real-time RT-PCR (right). B, The expression of IRF8 or IRF4 mRNAs in the cultured peritoneal macrophages (prepared from WT and Pdk2/4 DKO mice) following stimulation with LPS (100 ng/ml) plus IFN- (50 U/ml) or IL-4 (10 ng/ml) for 8 h was assessed by real-time RT-PCR. C, The expression of Pdk2 and Pdk4 mRNAs in the WT peritoneal macrophages following stimulation with LPS (100 ng/ml) plus IFN- (50 U/ml) for 8 h was assessed by real-time RT-PCR. Results for mRNA expression are displayed as the fold increase of gene expression normalized to GAPDH. *p 0.05 versus the control group. #p 0.05 between indicated groups (Student’s t test). n 3. Data are mean SEM.

Article Snippet: For immunofluorescence staining, sections were incubated with primary antibodies against PDK2 (rabbit, 1:200; Acris Antibodies), PDK4 (rabbit, 1:200; Atlas Antibodies), phospho-Ser 293-PDHE1 (pyruvate dehydrogenase E1 ) (rabbit, 1:200; Calbiochem), phospho-Ser 300-PDHE1 (pyruvate dehydrogenase E1 ) (rabbit, 1:200; Calbiochem), Iba-1 (rabbit, 1:1000; Wako; or goat, 1:200; Novus Biologicals [for costainings]), GFAP (mouse, 1:500; BD Biosciences), Ly6G (rat, 1:200; eBioscience) or iNOS (rabbit, 1:200; BD Biosciences), MYH1/2/3 (mouse, 1:200; Santa Cruz Biotechnology) and 3 tubulin (2G10) (mouse, 1:200; Santa Cruz Biotechnology), overnight at 4°C, and then incubated with FITC- or Cy3conjugated secondary antibodies (1:200; Jackson ImmunoResearch Laboratories).

Techniques: Cell Culture, Quantitative RT-PCR, Expressing, Gene Expression, Control

Figure 9. Pdk2 or Pdk4 single knock-out mice showed attenuated paw edema and pain responsestochronicinflammatoryinsult.A,CFAwasinjectedintotheplantarsurfaceoftheleft hindpaws (ipsilateral side) to induce local inflammation. Paw thicknesses were significantly greater in the ipsilateral sides than contralateral sides. Pdk2 or Pdk4 single-gene deficiency significantly reduced the CFA-induced increase in paw thickness compared with that of WT animals. PWT to force and PWL to heat were measured in contralateral and ipsilateral sides. In the ipsilateral sides, CFA injection reduced PWT to force (B) and PWL to heat (C). The chronic inflammation-inducedpainhypersensitivitieswereattenuatedinPdk2orPdk4single-geneKO micecomparedwithWTanimals.Nosignificantchangeinpain-relatedbehaviorwasobserved in the contralateral sides. *p 0.05 between ipsilateral sides of WT and Pdk2 KO mice. #p 0.05betweenipsilateralsidesofWTandPdk4KOmice(one-wayANOVAwithDunnett’sproce- dureforpawthicknessandPWL,Mann–WhitneytestforPWT).n 7.DataaremeanSEM.

Journal: Journal of Neuroscience

Article Title: Metabolic Connection of Inflammatory Pain: Pivotal Role of a Pyruvate Dehydrogenase Kinase-Pyruvate Dehydrogenase-Lactic Acid Axis

doi: 10.1523/jneurosci.1910-15.2015

Figure Lengend Snippet: Figure 9. Pdk2 or Pdk4 single knock-out mice showed attenuated paw edema and pain responsestochronicinflammatoryinsult.A,CFAwasinjectedintotheplantarsurfaceoftheleft hindpaws (ipsilateral side) to induce local inflammation. Paw thicknesses were significantly greater in the ipsilateral sides than contralateral sides. Pdk2 or Pdk4 single-gene deficiency significantly reduced the CFA-induced increase in paw thickness compared with that of WT animals. PWT to force and PWL to heat were measured in contralateral and ipsilateral sides. In the ipsilateral sides, CFA injection reduced PWT to force (B) and PWL to heat (C). The chronic inflammation-inducedpainhypersensitivitieswereattenuatedinPdk2orPdk4single-geneKO micecomparedwithWTanimals.Nosignificantchangeinpain-relatedbehaviorwasobserved in the contralateral sides. *p 0.05 between ipsilateral sides of WT and Pdk2 KO mice. #p 0.05betweenipsilateralsidesofWTandPdk4KOmice(one-wayANOVAwithDunnett’sproce- dureforpawthicknessandPWL,Mann–WhitneytestforPWT).n 7.DataaremeanSEM.

Article Snippet: For immunofluorescence staining, sections were incubated with primary antibodies against PDK2 (rabbit, 1:200; Acris Antibodies), PDK4 (rabbit, 1:200; Atlas Antibodies), phospho-Ser 293-PDHE1 (pyruvate dehydrogenase E1 ) (rabbit, 1:200; Calbiochem), phospho-Ser 300-PDHE1 (pyruvate dehydrogenase E1 ) (rabbit, 1:200; Calbiochem), Iba-1 (rabbit, 1:1000; Wako; or goat, 1:200; Novus Biologicals [for costainings]), GFAP (mouse, 1:500; BD Biosciences), Ly6G (rat, 1:200; eBioscience) or iNOS (rabbit, 1:200; BD Biosciences), MYH1/2/3 (mouse, 1:200; Santa Cruz Biotechnology) and 3 tubulin (2G10) (mouse, 1:200; Santa Cruz Biotechnology), overnight at 4°C, and then incubated with FITC- or Cy3conjugated secondary antibodies (1:200; Jackson ImmunoResearch Laboratories).

Techniques: Knock-Out, Injection

Figure 12. A proposed schematic outlining the implications of a PDK-PDH-lactic acid axis in the pathogenesis of chronic inflammatory pain. Inflammatory stimulus enhances the expression and activity of PDK2 and PDK4 at the site of inflammation, therebydecreasingtheoxidationofpyruvateandincreasingitsconversionintolactateviaphosphorylation/inhibitionofPDH.This metabolic shift-associated lactic acid production and resulting acidic microenvironment favor the recruitment of inflammatory cells to the site of inflammation and amplify the local inflammation ensuing the nociceptive responses. Enhanced PDK2/4 skew macrophagestowardtheM1(proinflammatory)phenotypeviaactivationofproinflammatoryphenotype-determiningtranscrip- tionfactorIRF8,resultingintheincreasedsecretionofproinflammatorymediators,suchasTNF-,IL-1,iNOS,andIL-6aswellas nitric oxide. The proalgesic mediators thus released activate nociceptors and spinal glia to cause peripheral and central sensitiza- tions,respectively.ThesefindingssuggestthatthePDK-PDH-lacticacidaxisplaysakeyroleinthepathogenesisofinflammation- induced chronic pain via the modulation of multifaceted proinflammatory events.

Journal: Journal of Neuroscience

Article Title: Metabolic Connection of Inflammatory Pain: Pivotal Role of a Pyruvate Dehydrogenase Kinase-Pyruvate Dehydrogenase-Lactic Acid Axis

doi: 10.1523/jneurosci.1910-15.2015

Figure Lengend Snippet: Figure 12. A proposed schematic outlining the implications of a PDK-PDH-lactic acid axis in the pathogenesis of chronic inflammatory pain. Inflammatory stimulus enhances the expression and activity of PDK2 and PDK4 at the site of inflammation, therebydecreasingtheoxidationofpyruvateandincreasingitsconversionintolactateviaphosphorylation/inhibitionofPDH.This metabolic shift-associated lactic acid production and resulting acidic microenvironment favor the recruitment of inflammatory cells to the site of inflammation and amplify the local inflammation ensuing the nociceptive responses. Enhanced PDK2/4 skew macrophagestowardtheM1(proinflammatory)phenotypeviaactivationofproinflammatoryphenotype-determiningtranscrip- tionfactorIRF8,resultingintheincreasedsecretionofproinflammatorymediators,suchasTNF-,IL-1,iNOS,andIL-6aswellas nitric oxide. The proalgesic mediators thus released activate nociceptors and spinal glia to cause peripheral and central sensitiza- tions,respectively.ThesefindingssuggestthatthePDK-PDH-lacticacidaxisplaysakeyroleinthepathogenesisofinflammation- induced chronic pain via the modulation of multifaceted proinflammatory events.

Article Snippet: For immunofluorescence staining, sections were incubated with primary antibodies against PDK2 (rabbit, 1:200; Acris Antibodies), PDK4 (rabbit, 1:200; Atlas Antibodies), phospho-Ser 293-PDHE1 (pyruvate dehydrogenase E1 ) (rabbit, 1:200; Calbiochem), phospho-Ser 300-PDHE1 (pyruvate dehydrogenase E1 ) (rabbit, 1:200; Calbiochem), Iba-1 (rabbit, 1:1000; Wako; or goat, 1:200; Novus Biologicals [for costainings]), GFAP (mouse, 1:500; BD Biosciences), Ly6G (rat, 1:200; eBioscience) or iNOS (rabbit, 1:200; BD Biosciences), MYH1/2/3 (mouse, 1:200; Santa Cruz Biotechnology) and 3 tubulin (2G10) (mouse, 1:200; Santa Cruz Biotechnology), overnight at 4°C, and then incubated with FITC- or Cy3conjugated secondary antibodies (1:200; Jackson ImmunoResearch Laboratories).

Techniques: Expressing, Activity Assay

Quantitative PCR primer sequences

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Defining the contribution of skeletal muscle pyruvate dehydrogenase α1 to exercise performance and insulin action

doi: 10.1152/ajpendo.00241.2018

Figure Lengend Snippet: Quantitative PCR primer sequences

Article Snippet: The following antibodies were used: PDHα (cat. no. 3205, Cell Signaling), eukaryotic translation elongation factor 2 (cat. no. 2332, Cell signaling), ATP synthase subunit alpha, ubiquinol-cytochrome C reductase core protein 2, mitochondrially encoded cytochrome C oxidase I, succinate dehydrogenase subunit B, NADH/ubiquinone oxidoreductase subunit B8 (cat. no. MS-604, MitoSciences), acyl-Coenzyme A dehydrogenase, very long-chain; cat. no. ab-155138, Abcam), acyl-Coenzyme A dehydrogenase, long-chain (cat. no. ab-82853, Abcam), hexokinase 2 (HK2; cat. no. 2857, Cell Signaling), lactate dehydrogenase A (cat. no. ABN-896, MilliporeSigma), Akt (cat. no. 2920, Cell Signaling), pAkt S473 (cat. no. 4058, Cell Signaling), GSK3α/β (5676, Cell signaling), pGSK3α/β S21/S9 (9331, Cell signaling), PDK4 (cat. no. NBP1-07047SS, Novus Biologicals), phosphofructokinase 1 (cat. no. sc-377346, Santa Cruz Biotechnology).

Techniques: Real-time Polymerase Chain Reaction

Metabolic and mitochondrial proteins in HFD-fed PDHmKO and WT mice. WT and PDHmKO mice were fed either a chow or a HFD for 12 wk. Transcript abundance of Pdha1, Pdk2, and Pdk4 (A) and Cd36, Cact (Slc25a20), and Acadl (B) in skeletal muscle from chow- or HFD-fed PDHmKO and WT mice, normalized to Ppib (Chow, n = 5/6; HFD, n = 6/7). Representative blot of PDHα, HK2, LDHA, ACADVL (C), and ATP5A, UQCRC2, MTCO1, SDHB, and NDUFB8 (D) protein abundance in skeletal muscle from chow- or HFD-fed PDHmKO and WT mice. Bar graphs show quantification of protein abundance in skeletal muscle relative to ponceau (Chow, n = 6/6; HFD, n = 6/6) (E). Data reported as means ± SE two-way ANOVA, #P < 0.05, main effect of diet, *P < 0.05, main effect of genotype (A–E). HFD, high-fat diet; PDHmKO, tamoxifen-inducible Pdha1 knockout mice; WT, wild-type.

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Defining the contribution of skeletal muscle pyruvate dehydrogenase α1 to exercise performance and insulin action

doi: 10.1152/ajpendo.00241.2018

Figure Lengend Snippet: Metabolic and mitochondrial proteins in HFD-fed PDHmKO and WT mice. WT and PDHmKO mice were fed either a chow or a HFD for 12 wk. Transcript abundance of Pdha1, Pdk2, and Pdk4 (A) and Cd36, Cact (Slc25a20), and Acadl (B) in skeletal muscle from chow- or HFD-fed PDHmKO and WT mice, normalized to Ppib (Chow, n = 5/6; HFD, n = 6/7). Representative blot of PDHα, HK2, LDHA, ACADVL (C), and ATP5A, UQCRC2, MTCO1, SDHB, and NDUFB8 (D) protein abundance in skeletal muscle from chow- or HFD-fed PDHmKO and WT mice. Bar graphs show quantification of protein abundance in skeletal muscle relative to ponceau (Chow, n = 6/6; HFD, n = 6/6) (E). Data reported as means ± SE two-way ANOVA, #P < 0.05, main effect of diet, *P < 0.05, main effect of genotype (A–E). HFD, high-fat diet; PDHmKO, tamoxifen-inducible Pdha1 knockout mice; WT, wild-type.

Article Snippet: The following antibodies were used: PDHα (cat. no. 3205, Cell Signaling), eukaryotic translation elongation factor 2 (cat. no. 2332, Cell signaling), ATP synthase subunit alpha, ubiquinol-cytochrome C reductase core protein 2, mitochondrially encoded cytochrome C oxidase I, succinate dehydrogenase subunit B, NADH/ubiquinone oxidoreductase subunit B8 (cat. no. MS-604, MitoSciences), acyl-Coenzyme A dehydrogenase, very long-chain; cat. no. ab-155138, Abcam), acyl-Coenzyme A dehydrogenase, long-chain (cat. no. ab-82853, Abcam), hexokinase 2 (HK2; cat. no. 2857, Cell Signaling), lactate dehydrogenase A (cat. no. ABN-896, MilliporeSigma), Akt (cat. no. 2920, Cell Signaling), pAkt S473 (cat. no. 4058, Cell Signaling), GSK3α/β (5676, Cell signaling), pGSK3α/β S21/S9 (9331, Cell signaling), PDK4 (cat. no. NBP1-07047SS, Novus Biologicals), phosphofructokinase 1 (cat. no. sc-377346, Santa Cruz Biotechnology).

Techniques: Quantitative Proteomics, Knock-Out

PDHmKO mice have reduced running speed during voluntary wheel running (VWR). Weekly average speed (km/h) (A), distance run per 24-h period (B), and time/24 h spent running (C) over 21 days VWR (n = 7/6). Representative blots of PDHα, HK2, PFK1, LDH, PDK4, ACADVL, ACADL, and NDUFB8 protein abundance in skeletal muscle from sedentary (SED) or exercise-trained (EXT) PDHmKO and WT mice (D). Bar graphs show quantification of protein abundance in skeletal muscle relative to ponceau (SED, n = 5/5; EXT, n = 7/6) (E). Data reported as means ± SE 2-way ANOVA, Sidak’s post hoc test, *P < 0.05, PDHmKO compared with WT (A–C); 2-way ANOVA, Tukey’s post hoc test, #P < 0.05, EXT compared with SED, *P < 0.05, PDHmKO compared with WT (D–E). PDHmKO, tamoxifen-inducible Pdha1 knockout mice; WT, wild-type.

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Defining the contribution of skeletal muscle pyruvate dehydrogenase α1 to exercise performance and insulin action

doi: 10.1152/ajpendo.00241.2018

Figure Lengend Snippet: PDHmKO mice have reduced running speed during voluntary wheel running (VWR). Weekly average speed (km/h) (A), distance run per 24-h period (B), and time/24 h spent running (C) over 21 days VWR (n = 7/6). Representative blots of PDHα, HK2, PFK1, LDH, PDK4, ACADVL, ACADL, and NDUFB8 protein abundance in skeletal muscle from sedentary (SED) or exercise-trained (EXT) PDHmKO and WT mice (D). Bar graphs show quantification of protein abundance in skeletal muscle relative to ponceau (SED, n = 5/5; EXT, n = 7/6) (E). Data reported as means ± SE 2-way ANOVA, Sidak’s post hoc test, *P < 0.05, PDHmKO compared with WT (A–C); 2-way ANOVA, Tukey’s post hoc test, #P < 0.05, EXT compared with SED, *P < 0.05, PDHmKO compared with WT (D–E). PDHmKO, tamoxifen-inducible Pdha1 knockout mice; WT, wild-type.

Article Snippet: The following antibodies were used: PDHα (cat. no. 3205, Cell Signaling), eukaryotic translation elongation factor 2 (cat. no. 2332, Cell signaling), ATP synthase subunit alpha, ubiquinol-cytochrome C reductase core protein 2, mitochondrially encoded cytochrome C oxidase I, succinate dehydrogenase subunit B, NADH/ubiquinone oxidoreductase subunit B8 (cat. no. MS-604, MitoSciences), acyl-Coenzyme A dehydrogenase, very long-chain; cat. no. ab-155138, Abcam), acyl-Coenzyme A dehydrogenase, long-chain (cat. no. ab-82853, Abcam), hexokinase 2 (HK2; cat. no. 2857, Cell Signaling), lactate dehydrogenase A (cat. no. ABN-896, MilliporeSigma), Akt (cat. no. 2920, Cell Signaling), pAkt S473 (cat. no. 4058, Cell Signaling), GSK3α/β (5676, Cell signaling), pGSK3α/β S21/S9 (9331, Cell signaling), PDK4 (cat. no. NBP1-07047SS, Novus Biologicals), phosphofructokinase 1 (cat. no. sc-377346, Santa Cruz Biotechnology).

Techniques: Quantitative Proteomics, Knock-Out