Journal: eLife

Article Title: Molecular mechanism for direct actin force-sensing by α-catenin

doi: 10.7554/eLife.62514

Figure Lengend Snippet:

Article Snippet: Gene ( H. sapiens ) , αE-catenin; Hs CTNNA1 , Addgene # 24194 , UniProt:P35221 , .

Techniques: Recombinant, Expressing, Plasmid Preparation, Synthesized, Transfection, Construct, Blocking Assay, Transmission Electron Microscopy, Software

Journal: Frontiers in Physiology

Article Title: Crocetin Attenuates Sepsis-Induced Cardiac Dysfunction via Regulation of Inflammatory Response and Mitochondrial Function

doi: 10.3389/fphys.2020.00514

Figure Lengend Snippet: Effect of CRO on mitochondrial function and morphology in LPS-induced cardiac sepsis. (A) Effect of CRO on mitochondrial respiratory function in LPS-induced H9c2 cells. Data ( n = 3) are presented as the mean ± SEM. (B,C) Effect of CRO on levels of ATP and BHB in LPS-induced H9c2 cells. Data ( n = 3) are presented as the mean ± SEM. *p < 0.05, *** p < 0.001. (D) Effect of CRO on the expression of ACAT1 and OXCT1 in LPS-induced H9c2 cells. Expression levels are summarized in panel (E) . Data ( n = 3) are presented as the mean ± SEM. *p < 0.05, ** p < 0.01, *** p < 0.001. (F) Effect of CRO (100 μM) on mitochondrial morphology in LPS-induced H9c2 cells. Mitochondrial length was determined in 50 individual cells using Leica software (G) . Data ( n = 3) are presented as the mean ± SEM, *p < 0.05, *** p < 0.001. (H) Effect of CRO on the expression of mitochondrial fusion and fission proteins in LPS-induced H9c2 cells. Expression levels are summarized in panel (I) . Data ( n = 3) are presented as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The membranes were blocked using 5% non-fat milk in TBST (Tris-buffered saline with 0.05% Tween 20) for 1 h, then incubated with primary antibodies against cytochrome c (1:1,000), Bcl-2 (1:500), Bax (1:1,000), caspase 3 (1:500), caspase 9 (1:500), PI3K (1:1,000), p-PI3K (1:1,000), p-Akt (1:1,000), Akt (1:1,000), p-p65 (1:1,000), p65 (1:1,000), NQO1 (1:1,000), Keap1 (1:1,000), Nrf2 (1:1,000), HO-1 (1:1,000), lamin B (1:1,000), ACAT1 (1:2,000), OXCT1 (1:2,000), Mfn1 (1:1,000), Mfn2 (1:2,000), OPA1 (1:1,000), Drp1 (1:2,000), Fis1 (1:2,000), MFF (1:1,000), Mid49 (1:1,000), and Mid51 (1:1,000, Cell Signaling Technology, United States) overnight at 4°C.

Techniques: Expressing, Software

Journal: Frontiers in Oncology

Article Title: Dysregulation of Ketone Body Metabolism Is Associated With Poor Prognosis for Clear Cell Renal Cell Carcinoma Patients

doi: 10.3389/fonc.2019.01422

Figure Lengend Snippet: mRNA expression of ACAT1, BDH2 , and HMGCL is downregulated in ccRCC in contrast to normal kidney samples . (A) A heatmap of the hierarchical clustering of 12 differentially expressed key genes involved in ketone body metabolism. Input data are the log2 ratios of mRNA expression values for ccRCCs over normal kidney tissues. Genes that are upregulated designated with red color, green color indicate downregulated genes. (B) The mRNA expression analysis of ACAT1, BDH2 , and HMGCL genes in 533 cases of ccRCC and 72 cases of normal kidney tissues based on TCGA database. (C) The mRNA expression analysis of ACAT1, BDH2 , and HMGCL genes by RT-qPCR in 14 primary renal cell carcinoma tissues and in the matched adjacent tissues, normalized to mRNA GAPDH a house-keeping gene expression level. *** p < 0.001.

Article Snippet: The full-length coding sequence for ACAT1, BDH2 , and HMGCL were subcloned into the pCMV6-Entry vector obtained from Origene (Rockville, MD, USA).

Techniques: Expressing, Quantitative RT-PCR

Journal: Frontiers in Oncology

Article Title: Dysregulation of Ketone Body Metabolism Is Associated With Poor Prognosis for Clear Cell Renal Cell Carcinoma Patients

doi: 10.3389/fonc.2019.01422

Figure Lengend Snippet: Detection of ACAT1, BDH2, and HMGCL protein expression in 85 cases of primary ccRCC tissues and matched adjacent kidney tissues. (A) Immunohistochemical staining. Representative slides of ccRCC and matched adjacent tissues samples were stained by ACAT1, BDH2, and HMGCL antibodies, (B) Analysis of IHC staining. Scores of IHC staining of all the investigated ccRCCs and corresponding adjacent non-cancerous stromal tissue samples. *** p < 0.001.

Article Snippet: The full-length coding sequence for ACAT1, BDH2 , and HMGCL were subcloned into the pCMV6-Entry vector obtained from Origene (Rockville, MD, USA).

Techniques: Expressing, Immunohistochemical staining, Staining, Immunohistochemistry

Journal: Frontiers in Oncology

Article Title: Dysregulation of Ketone Body Metabolism Is Associated With Poor Prognosis for Clear Cell Renal Cell Carcinoma Patients

doi: 10.3389/fonc.2019.01422

Figure Lengend Snippet: The expression of ACAT1, BDH2 , and HMGCL is significantly inter-correlated in ccRCCs. The pair-wise inter-correlation of transcriptional levels of ACAT1 and BDH2, ACAT1 and HMGCL, BDH2 and HMGCL based on TCGA database (A) and RT-qPCR data (B) . (C) The pair-wise inter-correlation of protein levels of ACAT1 and BDH2, ACAT1 and HMGCL, BDH2 and HMGCL based on IHC staining scores.

Article Snippet: The full-length coding sequence for ACAT1, BDH2 , and HMGCL were subcloned into the pCMV6-Entry vector obtained from Origene (Rockville, MD, USA).

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemistry

Journal: Frontiers in Oncology

Article Title: Dysregulation of Ketone Body Metabolism Is Associated With Poor Prognosis for Clear Cell Renal Cell Carcinoma Patients

doi: 10.3389/fonc.2019.01422

Figure Lengend Snippet: ROC curve for ACAT1, BDH2 , and HMGCL genes in ccRCC patients based on TCGA RNA-seq data. The ROC curves to evaluate the diagnostic value of ACAT1, BDH2 , and HMGCL expression, either independently in 533 cases of RCC and 72 cases of normal kidney tissues, or combining two genes or three genes.

Article Snippet: The full-length coding sequence for ACAT1, BDH2 , and HMGCL were subcloned into the pCMV6-Entry vector obtained from Origene (Rockville, MD, USA).

Techniques: RNA Sequencing Assay, Diagnostic Assay, Expressing

Journal: Frontiers in Oncology

Article Title: Dysregulation of Ketone Body Metabolism Is Associated With Poor Prognosis for Clear Cell Renal Cell Carcinoma Patients

doi: 10.3389/fonc.2019.01422

Figure Lengend Snippet: Association of ACAT1, BDH2 , and HMGCL expression with clinicopathological characteristics accordingly to TCGA mRNA-seq data.

Article Snippet: The full-length coding sequence for ACAT1, BDH2 , and HMGCL were subcloned into the pCMV6-Entry vector obtained from Origene (Rockville, MD, USA).

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: Dysregulation of Ketone Body Metabolism Is Associated With Poor Prognosis for Clear Cell Renal Cell Carcinoma Patients

doi: 10.3389/fonc.2019.01422

Figure Lengend Snippet: The lower expression of ACAT1, BDH2 , and HMGCL predicts the worse prognosis for ccRCC patients. Kaplan-Meier survival curves of overall survival (A,C,E) and disease-free survival (B,D,F) were plotted based on the mRNA level of ACAT1 , BDH2, and HMGCL , respectively. (G) Kaplan-Meier survival curves for ccRCC patients with combined values of ACAT1 , BDH2, and HMGCL expression based on TCGA mRNA-seq data.

Article Snippet: The full-length coding sequence for ACAT1, BDH2 , and HMGCL were subcloned into the pCMV6-Entry vector obtained from Origene (Rockville, MD, USA).

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: Dysregulation of Ketone Body Metabolism Is Associated With Poor Prognosis for Clear Cell Renal Cell Carcinoma Patients

doi: 10.3389/fonc.2019.01422

Figure Lengend Snippet: The tumor-suppressive effects of ACAT1, BDH2 , and HMGCL expression in ccRCC. (A) The mRNA levels of ACAT1, BDH2 , and HMGCL were determined by a real-time fluorescent quantitative PCR assay after transient transfection of 24 h. A bar chart shows a ratio of an mRNA expression value in cells transfected with a gene-coding vector to an mRNA value in cells transfected with an empty vector pCMV6. (B) The proliferation of 786-0 cells restored the expression of ACAT1, BDH2 , and HMGCL genes was measured by a CCK8 assay. (C) A transwell assay. The violet color dots represent cells penetrating through matrix gels. (D) Wound healing assay. Images were taken at 0 and 12 h after introducing a scratch in 786-0 cells. Gap closure was measured as mean ± SD of three independent experiments. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The full-length coding sequence for ACAT1, BDH2 , and HMGCL were subcloned into the pCMV6-Entry vector obtained from Origene (Rockville, MD, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Transfection, Plasmid Preparation, CCK-8 Assay, Transwell Assay, Wound Healing Assay

Journal: Frontiers in Oncology

Article Title: Dysregulation of Ketone Body Metabolism Is Associated With Poor Prognosis for Clear Cell Renal Cell Carcinoma Patients

doi: 10.3389/fonc.2019.01422

Figure Lengend Snippet: The effect of β-hydroxybutyrate on the proliferation and migration of 768-0 cells. The relative concentration of extracellular (A) and intracellular (B) β-hydroxybutyrate, when restoring the expression of ACAT1, BDH2, and HMGCL, respectively. (C) The proliferation of 786-0 cells was measured by a CCK8 assay upon the treatment of β-hydroxybutyrate with concentration of 0, 2.5, 5, and 10 mM. (D) Wound healing assay. Images were taken at 0 and 24 h after introducing a scratch in 786-0 cells. Gap closure was measured as mean ± SD of three independent experiments. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The full-length coding sequence for ACAT1, BDH2 , and HMGCL were subcloned into the pCMV6-Entry vector obtained from Origene (Rockville, MD, USA).

Techniques: Migration, Concentration Assay, Expressing, CCK-8 Assay, Wound Healing Assay