Journal: BMC musculoskeletal disorders

Article Title: Pyruvate Dehydrogenase Kinase 1 inhibition mediated oxidative phosphorylation enhancement in cartilage promotes osteoarthritis progression.

doi: 10.1186/s12891-023-06585-6

Figure Lengend Snippet: Fig. 2 PDK1 is involved in OA progression based on GEO. A Differential expression of PDK1 in GSE55235. B Differential expression of PDK1 in GSE98918. C The PPI network for PDK1 was constructed in GeneMANIA. D KEGG analysis for genes that highly correlated with PDK1. E GO analysis for genes that highly correlated with PDK1

Article Snippet: The sections were stained with a primary antibody against PDK1 (A01268-1, Boster), followed by the secondary antibody and Dapi.

Techniques: Quantitative Proteomics, Construct

Journal: BMC musculoskeletal disorders

Article Title: Pyruvate Dehydrogenase Kinase 1 inhibition mediated oxidative phosphorylation enhancement in cartilage promotes osteoarthritis progression.

doi: 10.1186/s12891-023-06585-6

Figure Lengend Snippet: Fig. 3 PDK1 is downregulated in human and mice osteoarthritic cartilage. A Immunofluorescence staining of PDK1 in articular cartilage from healthy and OA human. PDK1 stains red. Dapi stains the nucleus. B Quantification of an absolute number of PDK1.+ cells in articular cartilage from human. C Quantification analysis of the fluorescence intensity of PDK1 expression in articular cartilage from human. D Immunofluorescence staining of PDK1 in articular cartilage at sham or 4 weeks post-surgery. PDK1 stains red. Dapi stains the nucleus. E Quantifying an absolute number of PDK1 + cells in upper articular cartilage in the knee at the sham or 4 weeks post-surgery. F Quantification analysis of the fluorescence intensity of PDK1 expression in articular cartilage in the knee at sham or 4 weeks post-surgery. G PDH activity in articular cartilage at sham or 4 weeks post-surgery. In B, C, and E–G, horizontal lines and error bars show the mean ± SD (n ≥ 3 mice per group). * = P < 0.05; ** = P < 0.01; *** = P < 0.001

Article Snippet: The sections were stained with a primary antibody against PDK1 (A01268-1, Boster), followed by the secondary antibody and Dapi.

Techniques: Immunofluorescence, Staining, Fluorescence, Expressing, Activity Assay

Journal: BMC musculoskeletal disorders

Article Title: Pyruvate Dehydrogenase Kinase 1 inhibition mediated oxidative phosphorylation enhancement in cartilage promotes osteoarthritis progression.

doi: 10.1186/s12891-023-06585-6

Figure Lengend Snippet: Fig. 5 Suppressing of PDK1 disrupted anabolism and catabolism. A-C Representative immunohistochemistry images of Col2, Acan, MMP13 staining for mice at sham,4 or 8 weeks after DMM with or without jx06. D-F The mean ratio of integrated optical density (IOD) to area (IOD/area) was used to semi-quantify Col2, Acan, MMP13 amount, horizontal lines and error bars show the mean ± SD (n ≥ 3 mice per group). * = P < 0.05; ** = P < 0.01; *** = P < 0.001

Article Snippet: The sections were stained with a primary antibody against PDK1 (A01268-1, Boster), followed by the secondary antibody and Dapi.

Techniques: Immunohistochemistry, Staining

Journal: BMC musculoskeletal disorders

Article Title: Pyruvate Dehydrogenase Kinase 1 inhibition mediated oxidative phosphorylation enhancement in cartilage promotes osteoarthritis progression.

doi: 10.1186/s12891-023-06585-6

Figure Lengend Snippet: Fig. 6 Suppressing of PDK1 accelerated synovium inflammation. A Representative immunohistochemistry images of F4/80 staining for mice at sham,4 or 8 weeks after DMM with or without jx06. B Representative immunohistochemistry images of TNF-a staining for mice at sham,4 or 8 weeks after DMM with or without jx06. C Quantification of absolute number of F4/80.+ cells in synovium in the knee at sham or 4 weeks post-surgery. D The mean ratio of integrated optical density (IOD) to area (IOD/area) was used to semi-quantify TNF-a amount. E Model of PDK1 inhibitor modulates ECM degradation and synovium inflammation in mechanical stress-induced OA. Horizontal lines and error bars show the mean ± SD (n ≥ 3 mice per group). * = P < 0.05; ** = P < 0.01; *** = P < 0.001

Article Snippet: The sections were stained with a primary antibody against PDK1 (A01268-1, Boster), followed by the secondary antibody and Dapi.

Techniques: Immunohistochemistry, Staining

Journal: Scientific Reports

Article Title: The double inhibition of PDK1 and STAT3-Y705 prevents liver metastasis in colorectal cancer

doi: 10.1038/s41598-019-49480-8

Figure Lengend Snippet: High PDK1 expression is predictive of poor prognosis in CRC patients and promotes tumor growth in vivo . ( A ) Overall survival of CRC patients with high PDK1 expression (n = 42) was much shorter than patients with low PDK1 expression (n = 37). ( B ) IHC showed the representative results of low and high PDK1 expression based on the staining index on a tissue microarray. ( C ) Nude mice were subcutaneously injected with 8 × 10 6 HCT116 cells with or without the transduction of PDK1 shRNA. Silencing PDK1 significantly slowed down the growth of HCT116 xenografts ( P < 0.0001). ( D ) The gross xenografts were isolated from each nude mouse. ( E ) TUNEL assay showed that silencing PDK1 promoted the apoptosis of CRC cells in residual xenograft of nude mice. ( F ) Histogram displayed the corresponding comparison of the apoptosis cells per view presented in ( E ). Data expressed as mean ± S.D., **** represents P < 0.0001.

Article Snippet: The primary anti-PDK1 antibody ( cat#: 10026-1-AP , Proteintech Group , Inc , Rosemont , IL , USA ) was used at a dilution of 1:100.

Techniques: Expressing, In Vivo, Staining, Microarray, Injection, Transduction, shRNA, Isolation, TUNEL Assay, Comparison

Journal: Scientific Reports

Article Title: The double inhibition of PDK1 and STAT3-Y705 prevents liver metastasis in colorectal cancer

doi: 10.1038/s41598-019-49480-8

Figure Lengend Snippet: The direct interaction between PDK1 and p-STAT3 may contribute to CRC proliferation. ( A ) EdU incorporation assay showed silencing PDK1 obviously decreased the proliferation of HCT116 cells in vitro , compared with the control. ( B ) Histogram displayed the corresponding comparison of the proliferation rate presented in ( A ). ( C ) Colony formation assay demonstrated that knockdown of PDK1 significantly decreased HCT116 cell colony formation in vitro . ( D ) Histogram illustrated the corresponding comparison of colony formation numbers presented in ( C ). ( E ) Western blot showed that knockdown of PDK1 significantly reduced STAT3-p-Y705 protein level in HCT116 cells. In particular, the Y705 phosphorylation was completely inhibited by the combination of CPT (a STAT3-p-Y705 inhibitor) and knockdown of PDK1. Three gels were loaded, and blots from different proteins were cropped and grouped into one image with white area separated in between different proteins. The exposure time was 50 s, 30 s, 10 s for p-STAT3, STAT3 and GAPDH, respectively. ( F ) A Co-IP showed PDK1 interacted directly with STAT3 in both HCT116 cells and SW480 cells. Rabbit IgG was served as the control. One gel was loaded, and the blot for STAT3 protein was cropped (exposure time: 40 s). Data expressed as mean ± S.D., **** represents P < 0.0001.

Article Snippet: The primary anti-PDK1 antibody ( cat#: 10026-1-AP , Proteintech Group , Inc , Rosemont , IL , USA ) was used at a dilution of 1:100.

Techniques: In Vitro, Control, Comparison, Colony Assay, Knockdown, Western Blot, Phospho-proteomics, Co-Immunoprecipitation Assay

Journal: Scientific Reports

Article Title: The double inhibition of PDK1 and STAT3-Y705 prevents liver metastasis in colorectal cancer

doi: 10.1038/s41598-019-49480-8

Figure Lengend Snippet: Knockdown of PDK1 and inhibiting p-STAT3-Y705 decreased liver metastasis of colon cancer in the nude mice model. ( A ) 5 × 10 6 HCT116 cells with or without PDK1 shRNA transduction were injected into the spleens of nude mice under isoflurane anesthesia on day 1, respectively. On day 9, the nude mice were sacrificed, and the liver metastatic area was measured by ImageJ software. Compared with the control group, knockdown of PDK1 significantly inhibited liver metastasis of HCT116 cells. ( B ) SH-4-54 (an inhibitor of total STAT3, 0.2 mg/d/mouse) or CPT (0.2 mg/d/mouse) were injected (i.p.) every other day (day 1, 3, 5, 7) for a total of four times. The results showed that CPT plus knockdown of PDK1 markedly decreased the liver metastasis area, compared with the combination of SH-4-54 and knockdown of PDK1. ( C ) Histogram indicated the corresponding comparison of liver metastasis areas presented in ( A , B ). Data expressed as mean ± S.D., * represents P < 0.05, ** represents P < 0.01.

Article Snippet: The primary anti-PDK1 antibody ( cat#: 10026-1-AP , Proteintech Group , Inc , Rosemont , IL , USA ) was used at a dilution of 1:100.

Techniques: Knockdown, shRNA, Transduction, Injection, Software, Control, Comparison

Journal: Scientific Reports

Article Title: The double inhibition of PDK1 and STAT3-Y705 prevents liver metastasis in colorectal cancer

doi: 10.1038/s41598-019-49480-8

Figure Lengend Snippet: The combination of CPT and silencing PDK1 significantly inhibited liver metastasis of CRC by down-regulating p-STAT3-Y705 in the immune competent mice model. ( A ) 5 × 10 6 HCT116 cells with or without PDK1 shRNA transduction were injected into the spleens of BALB/C mice under isoflurane anesthesia, respectively. CPT (0.2 mg/d/mouse) were injected (i.p.) every other day (day 1, 3, 5, 7) for a total of four times. On day 9, the mice were sacrificed, and the liver metastatic area was measured by ImageJ software. The combination of CPT and PDK1 shRNA treatment led to a smallest liver metastasis area, compared with the control or PDK1 shRNA treatment alone. ( B ) Histogram indicated the corresponding comparison of liver metastasis areas presented in ( A ). Data expressed as mean ± S.D., **** represents P < 0.0001.

Article Snippet: The primary anti-PDK1 antibody ( cat#: 10026-1-AP , Proteintech Group , Inc , Rosemont , IL , USA ) was used at a dilution of 1:100.

Techniques: shRNA, Transduction, Injection, Software, Control, Comparison

Journal: Scientific Reports

Article Title: The double inhibition of PDK1 and STAT3-Y705 prevents liver metastasis in colorectal cancer

doi: 10.1038/s41598-019-49480-8

Figure Lengend Snippet: Silencing PDK1 significantly elevates the cytoplasmic ROS level. ( A ) HCT116 cells with or without the transduction of a PDK1 shRNA were cultured in normal condition. Flow cytometry showed ROS levels had no obvious difference regardless of PDK1 status. ( B ) As indicated in ( A ), cells were cultured in matrix detachment. ROS levels were significantly enhanced by silencing PDK1, compared with the controls. ( C ) Histogram indicated the corresponding comparison of ROS levels presented in normal and anoikis culture conditions in ( A , B ). Data expressed as mean ± S.D., **** represents P < 0.0001.

Article Snippet: The primary anti-PDK1 antibody ( cat#: 10026-1-AP , Proteintech Group , Inc , Rosemont , IL , USA ) was used at a dilution of 1:100.

Techniques: Transduction, shRNA, Cell Culture, Flow Cytometry, Comparison

Journal: Scientific Reports

Article Title: The double inhibition of PDK1 and STAT3-Y705 prevents liver metastasis in colorectal cancer

doi: 10.1038/s41598-019-49480-8

Figure Lengend Snippet: Anoikis partly contributes to the in vivo effect of the combination of knockdown of PDK1 and CPT. ( A ) HCT116 cells with or without the transduction of a PDK1 shRNA were seeded into 6-well plates and cultured in normal conditions. Then the cells were stained with AnnexinV-FITC/PI. Flow cytometry showed that the apoptosis rate was much higher after PDK1 was silenced. ( B ) As indicated in ( A ), the cells were seeded into 6-well plates coated with poly-HEMA. Flow cytometry showed that the apoptotic rate of HCT116 cells stably transduced with PDK1 was significantly higher than the other two controls. ( C ) Histogram indicated the corresponding comparison of the apoptosis rate presented in ( A , B ). ( D ) The results showed knockdown of PDK1 strongly sensitized CRC to anoikis; however, the anoikis rate was significantly reduced when PDK1 silencing was combined with CPT. ( E ) Histogram indicated the corresponding comparison of the apoptosis results presented in both normal condition and suspending condition in ( D ). Data expressed as mean ± S.D., *** represents P < 0.001, and **** represents P < 0.0001.

Article Snippet: The primary anti-PDK1 antibody ( cat#: 10026-1-AP , Proteintech Group , Inc , Rosemont , IL , USA ) was used at a dilution of 1:100.

Techniques: In Vivo, Knockdown, Transduction, shRNA, Cell Culture, Staining, Flow Cytometry, Stable Transfection, Comparison

Journal: Scientific Reports

Article Title: The double inhibition of PDK1 and STAT3-Y705 prevents liver metastasis in colorectal cancer

doi: 10.1038/s41598-019-49480-8

Figure Lengend Snippet: Knockdown of PDK1 plus CPT significantly reduces liver metastasis in CRC by downregulating the adherence capacity via inhibiting STAT3-p-Y705. ( A ) Transwell assay showed that the knockdown of PDK1 or the combination of CPT and silencing PDK1 significantly decreased migration capacity of liver metastatic HCT116 cells. ( B ) Histogram indicated the corresponding comparison of migration capacity presented in ( A ). ( C ) The liver metastatic HCT116 cells were seeded into a 6-well plate coated with fibronectin. The adherence assay showed that the knockdown of PDK1 significantly decreased the chemotaxis of liver metastatic HCT116 cells. In particular, the combination of CPT and knockdown of PDK1 resulted in a lowest cell adherence capacity. ( D ) Histogram indicated the corresponding comparison of the adherent cells presented in ( C ). Data expressed as mean ± S.D., * represents P < 0.05, ** represents P < 0.01, *** represents P < 0.001, and **** represents P < 0.0001.

Article Snippet: The primary anti-PDK1 antibody ( cat#: 10026-1-AP , Proteintech Group , Inc , Rosemont , IL , USA ) was used at a dilution of 1:100.

Techniques: Knockdown, Transwell Assay, Migration, Comparison, Chemotaxis Assay

Journal: Scientific Reports

Article Title: The double inhibition of PDK1 and STAT3-Y705 prevents liver metastasis in colorectal cancer

doi: 10.1038/s41598-019-49480-8

Figure Lengend Snippet: The potential mechanism of knockdown of PDK1 and CPT on inhibiting liver metastasis in CRC.

Article Snippet: The primary anti-PDK1 antibody ( cat#: 10026-1-AP , Proteintech Group , Inc , Rosemont , IL , USA ) was used at a dilution of 1:100.

Techniques: Knockdown

Journal: Cancer Communications

Article Title: Associations of PGK1 promoter hypomethylation and PGK1-mediated PDHK1 phosphorylation with cancer stage and prognosis: a TCGA pan-cancer analysis

doi: 10.1186/s40880-019-0401-9

Figure Lengend Snippet: PGK1 pS203 and PDHK1 pT338 levels increased from normal to tumor tissues in human cancers. PGK1 pS203 and PDHK1 pT338 levels in normal tissues, early-stage tumor tissues, and advanced tumor tissues were compared using immunohistochemical staining. Representative images are shown. a Breast carcinoma (BRCA). b Liver hepatocellular carcinoma (LIHC). c Lung adenocarcinoma (LUAD). d Stomach adenocarcinoma (STAD). e Esophageal carcinoma (ESCA). * P < 0.05, ** P < 0.01, *** P < 0.001 (2-tailed). PGK1 pS203 phosphorylated phosphoglycerate kinase 1 (PGK1) S203, PDHK1 pT338 phosphorylated phosphorylate pyruvate dehydrogenase kinase 1 (PDHK1) T338, HE hematoxylin–eosin staining

Article Snippet: After deparaffinization, rehydration, and antigen-retrieval, TMA slides were incubated with primary rabbit anti-human phospho-PGK1 S203 (dilution 1:200; Signalway Antibody; SAB487P), primary rabbit anti-human phospho-PDHK1 T338 (dilution 1:500; Signalway Antibody; #11596), or nonspecific IgG (as a negative control) overnight at 4 °C.

Techniques: Immunohistochemical staining, Staining

Journal: Cancer Communications

Article Title: Associations of PGK1 promoter hypomethylation and PGK1-mediated PDHK1 phosphorylation with cancer stage and prognosis: a TCGA pan-cancer analysis

doi: 10.1186/s40880-019-0401-9

Figure Lengend Snippet: PGK1 pS203 and PDHK1 pT338 levels are positively correlated with each other in human cancers. Scatter diagrams show the statistical results of the correlation between PGK1 pS203 and PDHK1 pT338 levels (analyzed using the Pearson correlation coefficient). The size of each dot reflects the number of specimens. PGK1 pS203 phosphorylated phosphoglycerate kinase 1 (PGK1) S203, PDHK1 pT338 phosphorylated phosphorylate pyruvate dehydrogenase kinase 1 (PDHK1) T338; BRCA breast carcinoma, LIHC liver hepatocellular carcinoma, LUAD lung adenocarcinoma, STAD stomach adenocarcinoma, ESCA esophageal carcinoma

Article Snippet: After deparaffinization, rehydration, and antigen-retrieval, TMA slides were incubated with primary rabbit anti-human phospho-PGK1 S203 (dilution 1:200; Signalway Antibody; SAB487P), primary rabbit anti-human phospho-PDHK1 T338 (dilution 1:500; Signalway Antibody; #11596), or nonspecific IgG (as a negative control) overnight at 4 °C.

Techniques:

Journal: Cancer Communications

Article Title: Associations of PGK1 promoter hypomethylation and PGK1-mediated PDHK1 phosphorylation with cancer stage and prognosis: a TCGA pan-cancer analysis

doi: 10.1186/s40880-019-0401-9

Figure Lengend Snippet: PGK1 pS203 and PDHK1 pT338 levels are associated with poor prognosis in cancer patients. K-Means cluster analysis was used to divide the indicated cancer patients into two groups with high and low levels of PGK1 pS203 and PDHK1 pT338. Kaplan–Meier survival curves were compared using the log-rank test. All statistical tests were two-sided. PGK1 pS203 phosphorylated phosphoglycerate kinase 1 (PGK1) S203, PDHK1 pT338 phosphorylated phosphorylate pyruvate dehydrogenase kinase 1 (PDHK1) T338, BRCA breast carcinoma, LIHC liver hepatocellular carcinoma, LUAD lung adenocarcinoma, STAD stomach adenocarcinoma, ESCA esophageal carcinoma

Article Snippet: After deparaffinization, rehydration, and antigen-retrieval, TMA slides were incubated with primary rabbit anti-human phospho-PGK1 S203 (dilution 1:200; Signalway Antibody; SAB487P), primary rabbit anti-human phospho-PDHK1 T338 (dilution 1:500; Signalway Antibody; #11596), or nonspecific IgG (as a negative control) overnight at 4 °C.

Techniques:

Journal: Cancer Communications

Article Title: Associations of PGK1 promoter hypomethylation and PGK1-mediated PDHK1 phosphorylation with cancer stage and prognosis: a TCGA pan-cancer analysis

doi: 10.1186/s40880-019-0401-9

Figure Lengend Snippet: Associations of PGK1 S203 and PDHK1 T338 phosphorylation levels with clinicopathologic characteristics in patients with BRCA, LIHC, LUAD, STAD and ESCA

Article Snippet: After deparaffinization, rehydration, and antigen-retrieval, TMA slides were incubated with primary rabbit anti-human phospho-PGK1 S203 (dilution 1:200; Signalway Antibody; SAB487P), primary rabbit anti-human phospho-PDHK1 T338 (dilution 1:500; Signalway Antibody; #11596), or nonspecific IgG (as a negative control) overnight at 4 °C.

Techniques: Phospho-proteomics, Immunohistochemistry

Journal: PLoS ONE

Article Title: Dichloroacetate affects proliferation but not apoptosis in canine mammary cell lines

doi: 10.1371/journal.pone.0178744

Figure Lengend Snippet: Western Blot analysis of PDK-1 expression in MTH53A, MTH52C, ZMTH3 and DT14/06T. All cell lines showed positivity for PDK-1 and GAPDH but no changes in PDK-1 expression was detectable between untreated and DCA exposed cells in any of the evaluated cell lines. GAPDH was used as loading control.

Article Snippet: The PVDF membrane was incubated overnight at 4°C with a canine specific polyclonal primary antibody anti-rabbit anti-PDK-1 (1:1000, ABIN2499649, antibodies-online, Aachen, Germany) or a canine specific polyclonal anti-rabbit anti-PDK-3 antibody (1:500, ABIN278250, antibodies-online).

Techniques: Western Blot, Expressing