Journal: Clinical and Translational Medicine

Article Title: The antineoplastic drug, trastuzumab, dysregulates metabolism in iPSC-derived cardiomyocytes

doi: 10.1186/s40169-016-0133-2

Figure Lengend Snippet: 76 genes were differentially expressed in both ERBB2-overexpressing cancer cell lines and iPSC-derived cardiomyocytes, following treatment with trastuzumab, p < 0.05

Article Snippet: For detection of PDK4 expression, blots were stained with 2 μg/mL dilution of anti-PDK4 (NBP1-07049, Novus Biologicals).

Techniques:

Journal: Clinical and Translational Medicine

Article Title: The antineoplastic drug, trastuzumab, dysregulates metabolism in iPSC-derived cardiomyocytes

doi: 10.1186/s40169-016-0133-2

Figure Lengend Snippet: Trastuzumab and lapatinib induced down-regulation of PHLDA1 protein expression and up-regulation of PDK4 protein expression. Western blot of 15 μg of total cell lysate of iPSC-derived cardiomyocytes stained for PHLDA1 and PDK4

Article Snippet: For detection of PDK4 expression, blots were stained with 2 μg/mL dilution of anti-PDK4 (NBP1-07049, Novus Biologicals).

Techniques: Expressing, Western Blot, Derivative Assay, Staining

Journal: Nature Communications

Article Title: FGF21 protects against HFpEF by improving cardiac mitochondrial bioenergetics in mice

doi: 10.1038/s41467-025-56885-9

Figure Lengend Snippet: A Volcano plot of proteomic analysis in AAV- Fgf21 - or AAV-GFP-treated mice with HFpEF. B Heatmap of the differentially expression proteins (DEPs) in the heart tissues of AAV- Fgf21 - or AAV-GFP-treated mice with HFpEF. C Relative expression levels of cardiac PDK4 in AAV- Fgf21 - or AAV-GFP-treated mice with HFpEF from the proteomic analysis data. n = 4 mice per condition. Cardiac PDK4 levels in Fgf21 KO and WT mice with or without HFpEF ( D ) or AAV- Fgf21 - or AAV-GFP-treated HFpEF mice ( E ), tested by immunoblot, respectively. n = 6 mice per condition. The expressional difference of cardiac PDK4 in the mitochondria ( F ) and non-mitochondria ( G ) of cardiomyocytes in HFpEF mice and controls was examined by immunoblot, respectively. n = 6 mice per condition. Data are presented as mean ± SEM. Statistical significance was determined by the two-tailed unpaired Student’s t test ( A , F , G ), two-sided Brown-Forsythe and Welch ANOVA with Dunnett’s multiple comparison test ( C ), or two-way ANOVA with Tukey’s multiple comparison test ( D , E ).

Article Snippet: AAV9 expressing PDK4 was constructed and produced by Shanghai OBiO Technology Co., Ltd.

Techniques: Expressing, Western Blot, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: FGF21 protects against HFpEF by improving cardiac mitochondrial bioenergetics in mice

doi: 10.1038/s41467-025-56885-9

Figure Lengend Snippet: A Average systolic blood pressure (SBP, left) and diastolic blood pressure (DBP, right) of Fgf21 KO and WT mice with HFpEF, after treatment with or without Pdk4 shRNA. n = 6 mice per condition. B The body weight of mice mentioned above. n = 6 mice per condition. C Left ventricular diastolic function of mice mentioned above. Top, pulsed-wave Doppler (top) and tissue Doppler (bottom) tracings. Bottom, quantification of the ratio between the mitral E wave and A wave (E/A), and the mitral E wave and E′ wave (E/E′). n = 6 mice per condition. D The lung weight to tibia length ratio (LW/TL) of mice mentioned above. n = 6 mice per condition. Running distance during exercise exhaustion test ( E ) and the heart weight to tibia length ratio (HW/TL, F ) of mice mentioned above. n = 6 mice per condition. G Hematoxylin and eosin (H&E) and wheat germ agglutinin (WGA) staining of cardiac sections (left) and cardiomyocyte cross-sectional area quantification (right) in mice mentioned above. Scale bars, 1 mm (H&E), and 25 μm (WGA). n = 50 cardiomyocytes per condition. H The mRNA expression levels of cardiac hypertrophic marker genes ( Anp , Bnp , and Myh7 ) in mice mentioned above. n = 6 mice per condition. I Dihydroethidium (DHE) staining (left) of heart tissues and quantification (right) of the intensity, Scale bars, 100 μm. n = 6 mice per condition. J Sirius red staining of heart tissues (left) and quantification of the fibrotic areas (right). Scale bars, 50 μm. n = 6 mice per condition. K Cardiac contents of fibrotic factors, including fibronectin, collagen I, and α-SMA tested by immunoblot. n = 6 mice per condition. L Cardiac p-PDH, PDH and PDK4 contents of mice tested by immunoblot. n = 6 mice per condition. M Cardiac pyruvate dehydrogenase activity in mice mentioned above. n = 6 mice per condition. Cardiac pyruvate ( N ), acetyl-CoA ( O ), and ATP ( P ) contents in mice mentioned above. n = 6 mice per condition. Violin plots in ( G ) are presented as lines indicating the median and interquartile range; other bar graphs are presented as mean ± SEM. Statistical significance was determined by the two-way ANOVA, followed by Tukey’s multiple comparison test.

Article Snippet: AAV9 expressing PDK4 was constructed and produced by Shanghai OBiO Technology Co., Ltd.

Techniques: shRNA, Staining, Expressing, Marker, Western Blot, Activity Assay, Comparison

Journal: Nature Communications

Article Title: FGF21 protects against HFpEF by improving cardiac mitochondrial bioenergetics in mice

doi: 10.1038/s41467-025-56885-9

Figure Lengend Snippet: A Cardiac p-PI3K, PI3K, p-AKT, AKT levels in Fgf21 KO and WT mice with or without HFpEF. n = 6 mice per condition. B Cardiac p-PI3K, PI3K, p-AKT, AKT contents in mice with HFpEF, treated with AAV- Fgf21 or AAV-GFP. n = 6 mice per condition. C – I Neonatal rat ventricular myocytes (NRVMs) were incubated with palmitic acid (PA) before administration of recombinant mouse FGF21 (rmFGF21), and then treated with LY294002 or vehicle for 24 h. For Flag- Pdk4 transfection, NRVMs were infected with Flag- Pdk4 for 48 h, then incubated with PA and rmFGF21, followed by treatment with LY294002 or vehicle. C PDK4, p-PDH, and PDH levels in NRVMs were tested by immunoblot. n = 5 biological samples. The activity of pyruvate dehydrogenase ( D ), pyruvate ( E ), acetyl-CoA ( F ), and ATP ( G ) contents in NRVMs. n = 5 biological samples. H Representative images of DCFH-DA staining and MitoSOX Green staining (top) and quantification of the DCFH-DA intensity and mitochondrial length (bottom). Scale bars, 50 μm for DCFH-DA staining, 10 μm (top) and 2 μm (bottom) for MitoSOX Green staining. n = 5 biological samples. I Representative images of JC-1 staining (left) and quantification of the red/green fluorescence intensity ratio (right). Scale bars, 50 μm. Aggregates, red; Monomers, green. n = 5 biological samples. Data are presented as mean ± SEM. Statistical significance was determined by two-way ANOVA, followed by Tukey’s multiple comparison test ( A , B ). Other assays were assessed by one-way ANOVA, followed by Tukey’s multiple comparison test.

Article Snippet: AAV9 expressing PDK4 was constructed and produced by Shanghai OBiO Technology Co., Ltd.

Techniques: Incubation, Recombinant, Transfection, Infection, Western Blot, Activity Assay, Staining, Fluorescence, Comparison

Journal: Nature Communications

Article Title: FGF21 protects against HFpEF by improving cardiac mitochondrial bioenergetics in mice

doi: 10.1038/s41467-025-56885-9

Figure Lengend Snippet: A Average systolic blood pressure (SBP, left) and diastolic blood pressure (DBP, right) of Apn KO and WT mice with HFpEF after treatment with AAV- Fgf21 or AAV-GFP, respectively. n = 6 mice per condition. B The body weight of mice mentioned above. n = 6 mice per condition. C Left ventricular diastolic function of Apn KO and WT mice with HFpEF, treated with AAV- Fgf21 or AAV-GFP, respectively. Top, representative pulsed-wave Doppler (top) and tissue Doppler (bottom) tracings of mice. Bottom, quantification of the ratio between the mitral E wave and A wave (E/A), and the mitral E wave and E′ wave (E/E′). n = 6 mice per condition. D The lung weight to tibia length ratio (LW/TL) of mice mentioned above. n = 6 mice per condition. Running distance during exercise exhaustion test ( E ) and the heart weight to tibia length ratio (HW/TL, F ) of mice mentioned above. n = 6 mice per condition. G Hematoxylin and eosin (H&E) and wheat germ agglutinin (WGA) staining of cardiac sections (left) and cardiomyocyte cross-sectional area quantification (right) in mice mentioned above. Scale bars, 1 mm (H&E), and 50 μm (WGA). n = 50 cardiomyocytes per condition. H The mRNA expression levels of cardiac hypertrophic marker genes ( Anp , Bnp , and Myh7 ) in mice mentioned above. n = 6 mice per condition. I Dihydroethidium (DHE) staining (left) of heart tissues and quantification (right) of the intensity, Scale bars, 100 μm. n = 6 mice per condition. J Sirius red staining of heart tissues (left) and quantification of the fibrotic areas (right). Scale bars, 50 μm. n = 6 mice per condition. K Cardiac contents of fibrotic factors, including fibronectin, collagen I, and α-SMA tested by immunoblot. n = 6 mice per condition. L Cardiac PDK4, p-PDH and PDH contents of mice tested by immunoblot. n = 6 mice per condition. M Cardiac pyruvate dehydrogenase activity in mice mentioned above. n = 6 mice per condition. Cardiac pyruvate ( N ), acetyl-CoA ( O ), and ATP ( P ) contents in mice mentioned above. n = 6 mice per condition. Violin plots in ( G ) are presented as lines indicating the median and interquartile range; other bar graphs are presented as mean ± SEM. Statistical significance was assessed by two-way ANOVA, followed by Tukey’s multiple comparison test.

Article Snippet: AAV9 expressing PDK4 was constructed and produced by Shanghai OBiO Technology Co., Ltd.

Techniques: Staining, Expressing, Marker, Western Blot, Activity Assay, Comparison

Journal: Nature Communications

Article Title: FGF21 protects against HFpEF by improving cardiac mitochondrial bioenergetics in mice

doi: 10.1038/s41467-025-56885-9

Figure Lengend Snippet: Neonatal rat ventricular myocytes (NRVMs) were co-cultured with subcutaneous adipose tissues isolated from WT (wt-SAT) or Apn KO (ko-SAT) mice for 24 h after 48 h of Flag- Pdk4 transfection, then treated with palmitic acid (PA), followed by incubation with recombinant mouse FGF21 (rmFGF21) or rmFGF21 plus LY294002 for 24 h. A p-PI3K, PI3K, p-AKT, AKT, PDK4, p-PDH, and PDH levels in NRVMs were tested by immunoblot. n = 4 biological samples. The activity of pyruvate dehydrogenase ( B ), pyruvate ( C ), acetyl-CoA ( D ), and ATP ( E ) contents in NRVMs. n = 5 biological samples. F Representative images of DCFH-DA staining and MitoSOX Green staining (top) and quantification of the DCFH-DA intensity and mitochondrial length (bottom). Scale bars, 50 μm for DCFH-DA staining, 10 μm (top) and 2 μm (bottom) for MitoSOX Green staining. n = 5 biological samples. G Representative images of JC-1 staining (left) and quantification of the red/green fluorescence intensity ratio (right). Scale bars, 50 μm. Aggregates, red; Monomers, green. n = 5 biological samples. H Real-time monitoring of the oxygen consumption rate (OCR) in NRVMs. n = 5 biological samples. Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA, followed by Tukey’s multiple comparison test.

Article Snippet: AAV9 expressing PDK4 was constructed and produced by Shanghai OBiO Technology Co., Ltd.

Techniques: Cell Culture, Isolation, Transfection, Incubation, Recombinant, Western Blot, Activity Assay, Staining, Fluorescence, Comparison

Journal: Molecular Oncology

Article Title: Reversing tozasertib resistance in glioma through inhibition of pyruvate dehydrogenase kinases

doi: 10.1002/1878-0261.13025

Figure Lengend Snippet: Transcriptome profiling analysis reveals that PDK4 is involved in tozasertib resistance in human malignant glioma cell lines. (A) Schematic of the approach taken to identify tozasertib‐responsive genes. (B, C) Heat map representation of the top 40 DEGs (based on log 2 fold change and adjusted P value < 0.05) comparing drug‐naïve, tozasertib‐resistant (resistant), or cells treated with 100 nmol·L −1 tozasertib for 3 days (short‐term) in U87 cells (B), and LNZ308 cells (C). (D) Venn diagram indicating number of genes overlapping in U87‐resistant and LNZ308‐resistant cells between comparisons. (E) KEGG pathway enrichment plot comparing resistant to naïve LNZ308 cells. Each circle in the figure represents a pathway further split into activated and suppressed. The Y‐axis represents the name of the pathway, and the X ‐axis indicates Enrichment Factor, indicating the proportion of the annotated genes to all genes in the pathway. (F) PDK4 mRNA expression across several glioma tumors compared with normal brain in the REMBRANDT dataset. Meta‐analysis performed in R2 genomics software comparing normal brain (cortex, n = 28), astrocytoma ( n = 147), oligodendroglioma ( n = 67), and GBM ( n = 219) showed PDK4 mRNA expression was significantly upregulated compared to normal brain (** P < 0.001 normal brain vs astrocytoma; * P < 0.05 normal brain vs oligodendroglioma; * P < 0.01 normal brain vs GBM).

Article Snippet: For transient transfection, commercially available ON‐TARGET‐plus siRNAs for human aurora kinase A (siRNA‐1, J‐003545‐26‐0002 and siRNA‐2, J‐003545‐27‐0002); aurora kinase B (siRNA‐1, J‐003326‐21‐0002 and siRNA‐2, J‐003326‐22‐0002); aurora kinase C (siRNA‐1, J‐019573‐11‐0002 and siRNA‐2, J‐019573‐13‐0002); and non‐target control siRNA (NT‐siRNA, D‐001830‐01‐05) sequences were used and transfected following the manufacturer's protocol (Dharmacon, Lafayette, CO, USA). pCMV‐6 vector (catalog number PS100001) and PDK4 (catalog number CW306843) expression plasmids obtained from Origene (Rockville, MD, USA) were used for overexpression analysis as reported previously [ , ].

Techniques: Expressing, Software

Journal: Molecular Oncology

Article Title: Reversing tozasertib resistance in glioma through inhibition of pyruvate dehydrogenase kinases

doi: 10.1002/1878-0261.13025

Figure Lengend Snippet: Sustained exposure to tozasertib promotes PDK expression and inhibits PDH activity. (A) Whole cell extracts from drug‐naïve, or cells treated with tozasertib for 72 h (short‐term), or tozasertib‐resistant cells were prepared. Twenty micrograms of protein were separated on a SDS/PAGE. Western blot analysis was performed with the indicated antibodies. Expression of β‐actin served as a loading control. These experiments were performed at least three times, and a representative blot is presented here. (B) Whole cell extracts from drug‐naïve or tozasertib‐resistant cells were subjected to PDH activity assay and reported as PDH activity·min −1 ·mL −1 . The values are represented as mean ± standard deviation of three independent experiments (statistical analysis was carried out with an unpaired two‐tailed t ‐test; ** P < 0.005, *** P < 0.0001). (C–E) Drug‐naïve or tozasertib‐resistant adult (U87) and pediatric (SJG2 and SF8628) glioma cell lines were treated with 10 µmol·L −1 of PDK4‐IN‐1 hydrochloride (C) or 10 µmol·L −1 of VER‐246608 (D) or 1 mmol·L −1 DCA (E) for 72 h. Vehicle (DMSO) treated cells served as control. Seventy‐two hours later, cell growth was assessed by MTS assay. Absorbance was recorded at 490 nm using a Synergy HTX multimode microplate reader. The values are represented as mean ± standard deviation of three separate experiments (= four wells per condition). Data (C–E) were analyzed for statistical significance by Tukey's ANOVA (NS, not significant, * P < 0.01, *** P < 0.0005). (F, G) Logarithmically growing U87, SJG2, and SF8628 drug naïve or tozasertib‐resistant cells were transfected with empty vector or PDK4 expression plasmid. Forty‐eight hours after infection, cells were washed, lifted, and lysed. Equal amount of protein was separated on a SDS/PAGE and western blot analysis was performed with PDK4 antibody. Expression of β‐actin served as a loading control. These experiments were performed at least three times, and a representative blot (F, upper panel) is presented here. In parallel, cell lysates were analyzed for PDH activity and reported as fold change relative to control (F, lower panel). Data are the mean ± SD of three independent experiments (*** P < 0.0005, data analyzed for statistical significance by unpaired two‐tailed t ‐test). (G) Vector or PDK4‐transfected cells were treated with or without tozasertib (100 nmol·L −1 ) for 72 h. Vehicle (DMSO) treated cells served as control. Cell growth was assessed by spectrophotometric measurement of MTS bioreduction using a microplate reader ( n = 4 wells per condition). Data point represents the mean ± SD of three measurements. Statistical analysis was carried out with Tukey's ANOVA (** P < 0.001, *** P < 0.0001).

Article Snippet: For transient transfection, commercially available ON‐TARGET‐plus siRNAs for human aurora kinase A (siRNA‐1, J‐003545‐26‐0002 and siRNA‐2, J‐003545‐27‐0002); aurora kinase B (siRNA‐1, J‐003326‐21‐0002 and siRNA‐2, J‐003326‐22‐0002); aurora kinase C (siRNA‐1, J‐019573‐11‐0002 and siRNA‐2, J‐019573‐13‐0002); and non‐target control siRNA (NT‐siRNA, D‐001830‐01‐05) sequences were used and transfected following the manufacturer's protocol (Dharmacon, Lafayette, CO, USA). pCMV‐6 vector (catalog number PS100001) and PDK4 (catalog number CW306843) expression plasmids obtained from Origene (Rockville, MD, USA) were used for overexpression analysis as reported previously [ , ].

Techniques: Expressing, Activity Assay, SDS Page, Western Blot, Control, Standard Deviation, Two Tailed Test, MTS Assay, Transfection, Plasmid Preparation, Infection

Journal: Scientific Reports

Article Title: Down-regulation of PDK4 is Critical for the Switch of Carbohydrate Catabolism during Syncytialization of Human Placental Trophoblasts

doi: 10.1038/s41598-017-09163-8

Figure Lengend Snippet: PDK4 expression during syncytialization of human placental trophoblasts. ( A ) Changes of the RPKM values for PDK family members in trophoblasts before (red column, 3 hours) and after (blue column, 48 hours) syncytialization. ( B ) Changes in PDK4 mRNA (black column, n = 4) and protein (white column, n = 4) abundance during syncytialization. ( C ) Changes in the phosphorylation of PDHE1α during syncytialization n = 4. ( D ) Representative images showing intense staining of PDK4 (red) in the cytotrophoblast layer and weak staining of PDK4 in the syncytial layer of human chorionic villi at early gestation. The syncytial and cytotrophoblast layers were labeled with β-hCG (green) and SPINT1 (green) respectively. Nuclei were counterstained with DAPI (blue). n = 3; *P < 0.05; **P < 0.01; ***P < 0.001 against 3 hours; n.s., not significant.

Article Snippet: To study the roles of PDK4 in the regulation of carbohydrate metabolism during syncytialization, the cells were transfected with small interfering RNA (siRNA) targeting PDK4 (sense: GAUGCUCUGUGAUCAGUAUTT, antisense: AUACUGAUCACAGAGCAUCTT) (GenePharma Co., Ltd., Shanghai, China) or a eukaryotic vector GV230 expressing PDK4 (GeneChem Co., Ltd. Shanghai, China) immediately after isolation.

Techniques: Expressing, Staining, Labeling

Journal: Scientific Reports

Article Title: Down-regulation of PDK4 is Critical for the Switch of Carbohydrate Catabolism during Syncytialization of Human Placental Trophoblasts

doi: 10.1038/s41598-017-09163-8

Figure Lengend Snippet: Role of PDK4 in the switch of carbohydrate catabolism in human placental trophoblasts. ( A ) siRNA-mediated knock-down of PDK4 (siPDK4) for 48 hours significantly decreased PDK4 mRNA abundance. n = 6. ( B ) Knock-down of PDK4 (siPDK4) for 48 hours significantly reduced PDHE1α phosphorylation. n = 3. ( C) and ( D ) siRNA-mediated knock-down of PDK4 decreased cellular lactate levels (n = 5, C ) while increased cellular ATP levels (n = 5, D ). Randomly scrambled siRNA served as negative control (Nc). (E) Over-expression of PDK4 by transfecting PDK4 plasmid significantly improved PDK4 mRNA abundance. n = 3. (F) Over-expression of PDK4 significantly increased PDHE1α phosphorylation. n = 3. ( G ) and (H ) Over-expression of PDK4 increased lactate levels (n = 5, G ) while decreased cellular ATP levels (n = 5, H ). *P < 0.05; **P < 0.01 against Nc ( A to D ) or Vehicle ( E to H ).

Article Snippet: To study the roles of PDK4 in the regulation of carbohydrate metabolism during syncytialization, the cells were transfected with small interfering RNA (siRNA) targeting PDK4 (sense: GAUGCUCUGUGAUCAGUAUTT, antisense: AUACUGAUCACAGAGCAUCTT) (GenePharma Co., Ltd., Shanghai, China) or a eukaryotic vector GV230 expressing PDK4 (GeneChem Co., Ltd. Shanghai, China) immediately after isolation.

Techniques: Negative Control, Over Expression, Plasmid Preparation

Journal: Scientific Reports

Article Title: Down-regulation of PDK4 is Critical for the Switch of Carbohydrate Catabolism during Syncytialization of Human Placental Trophoblasts

doi: 10.1038/s41598-017-09163-8

Figure Lengend Snippet: Role of PDK4 in syncytiotrophoblast functions. ( A ) and ( B ) The abundance of 11β-HSD2 mRNA (n = 4, A ) and protein (n = 3, B ) and β-hCG mRNA (n = 4, A ) was significantly increased by siRNA-mediated knock-down of PDK4 expression. Randomly scrambled siRNA served as negative control (Nc). (C) and ( D ) Over-expression of PDK4 significantly decreased the abundance of 11β-HSD2 mRNA (n = 4, C ) and protein (n = 3, D ) and β-hCG mRNA (n = 4, C ). *P < 0.05; **P < 0.01; ***P < 0.001 against Nc ( A and B ) or Vehicle ( C and D ).

Article Snippet: To study the roles of PDK4 in the regulation of carbohydrate metabolism during syncytialization, the cells were transfected with small interfering RNA (siRNA) targeting PDK4 (sense: GAUGCUCUGUGAUCAGUAUTT, antisense: AUACUGAUCACAGAGCAUCTT) (GenePharma Co., Ltd., Shanghai, China) or a eukaryotic vector GV230 expressing PDK4 (GeneChem Co., Ltd. Shanghai, China) immediately after isolation.

Techniques: Expressing, Negative Control, Over Expression

Journal: Scientific Reports

Article Title: Down-regulation of PDK4 is Critical for the Switch of Carbohydrate Catabolism during Syncytialization of Human Placental Trophoblasts

doi: 10.1038/s41598-017-09163-8

Figure Lengend Snippet: Role of activation of the cAMP/PKA signaling pathway by hCG in the regulation of PDK4 expression in human placental trophoblasts. ( A ) and ( B ) Treatment of trophoblasts with dibutyryl-cAMP (db-cAMP, 200 μM) for 24 hours significantly decreased the amounts of PDK4 mRNA (n = 6, A ) and protein (n = 3, B ). ( C ) and ( D ) Treatment of trophoblasts with β-hCG antibody (hCGab, 1:100) or PKA inhibitor H89 (20 μM) for 24 hours significantly increased the abundance of PDK4 mRNA (n = 4, C ) and protein (n = 3, D ). *P < 0.05; **P < 0.01 against Ctr.

Article Snippet: To study the roles of PDK4 in the regulation of carbohydrate metabolism during syncytialization, the cells were transfected with small interfering RNA (siRNA) targeting PDK4 (sense: GAUGCUCUGUGAUCAGUAUTT, antisense: AUACUGAUCACAGAGCAUCTT) (GenePharma Co., Ltd., Shanghai, China) or a eukaryotic vector GV230 expressing PDK4 (GeneChem Co., Ltd. Shanghai, China) immediately after isolation.

Techniques: Activation Assay, Expressing

Journal: Scientific Reports

Article Title: Down-regulation of PDK4 is Critical for the Switch of Carbohydrate Catabolism during Syncytialization of Human Placental Trophoblasts

doi: 10.1038/s41598-017-09163-8

Figure Lengend Snippet: Diagram illustrating the switch from glycolysis to OXPHOS via down-regulation of PDK4 expression upon activation of the cAMP pathway by hCG during syncytialization of the trophoblasts.

Article Snippet: To study the roles of PDK4 in the regulation of carbohydrate metabolism during syncytialization, the cells were transfected with small interfering RNA (siRNA) targeting PDK4 (sense: GAUGCUCUGUGAUCAGUAUTT, antisense: AUACUGAUCACAGAGCAUCTT) (GenePharma Co., Ltd., Shanghai, China) or a eukaryotic vector GV230 expressing PDK4 (GeneChem Co., Ltd. Shanghai, China) immediately after isolation.

Techniques: Expressing, Activation Assay

Journal: Scientific Reports

Article Title: Down-regulation of PDK4 is Critical for the Switch of Carbohydrate Catabolism during Syncytialization of Human Placental Trophoblasts

doi: 10.1038/s41598-017-09163-8

Figure Lengend Snippet: Information of the antibodies for immunohistochemical staining and western blotting.

Article Snippet: To study the roles of PDK4 in the regulation of carbohydrate metabolism during syncytialization, the cells were transfected with small interfering RNA (siRNA) targeting PDK4 (sense: GAUGCUCUGUGAUCAGUAUTT, antisense: AUACUGAUCACAGAGCAUCTT) (GenePharma Co., Ltd., Shanghai, China) or a eukaryotic vector GV230 expressing PDK4 (GeneChem Co., Ltd. Shanghai, China) immediately after isolation.

Techniques: Immunohistochemical staining, Staining, Western Blot

Journal: Scientific Reports

Article Title: Down-regulation of PDK4 is Critical for the Switch of Carbohydrate Catabolism during Syncytialization of Human Placental Trophoblasts

doi: 10.1038/s41598-017-09163-8

Figure Lengend Snippet: Primer sequences for qRT-PCR.

Article Snippet: To study the roles of PDK4 in the regulation of carbohydrate metabolism during syncytialization, the cells were transfected with small interfering RNA (siRNA) targeting PDK4 (sense: GAUGCUCUGUGAUCAGUAUTT, antisense: AUACUGAUCACAGAGCAUCTT) (GenePharma Co., Ltd., Shanghai, China) or a eukaryotic vector GV230 expressing PDK4 (GeneChem Co., Ltd. Shanghai, China) immediately after isolation.

Techniques:

Journal: Cell Death & Disease

Article Title: Loss of the novel mitochondrial protein FAM210B promotes metastasis via PDK4-dependent metabolic reprogramming

doi: 10.1038/cddis.2017.273

Figure Lengend Snippet: FAM210B knockdown cells show decreased PDK4 expression. ( a ) Heat map of the top 25 differentially regulated genes in siFAM210B and negative control A549 cells. ( b ) Quantitative PCR analyses of the relative expression of the indicated genes in siFAM210B normalized to negative control SKOV3 cells. ( c ) Immunoblotting for PDK4 in the indicated treated SKOV3 cells. ( d ) Schematically depicted PDK function in SKOV3 cell metabolism. ( e ) Western blotting analysis of the phosphorylation levels of PDH-E1 α in the indicated treated SKOV3 cells. ( f ) Normalized OCR and ( g ) normalized ECAR in siPDK4 ( n =6) and negative control SKOV3 cells ( n =8 wells)

Article Snippet: For the overexpression of PDK4, recombinant adenovirus expressing PDK4 and empty control adenovirus were constructed by Shandong Vigene Biosciences.

Techniques: Knockdown, Expressing, Negative Control, Real-time Polymerase Chain Reaction, Western Blot, Phospho-proteomics