Journal: International Journal of Molecular Sciences

Article Title: Role of Clostridium perfringens Enterotoxin on YAP Activation in Colonic Sessile Serrated Adenoma/Polyps with Dysplasia

doi: 10.3390/ijms21113840

Figure Lengend Snippet: Effect of CPE on EMT phenotype in HT29 and HCM116 human colon cancer cells. ( A ) Effect of CPE on cell growth in the two cell lines. ( B , C ) Effect of CPE pretreatment on cell growth. HT29 cells ( B ) and HCT116 cells were treated with CPE of IC5 for 24 h. ( C ). ( D ) Effect of CPE on cell invasion. ( E ) Effect of CPE on protein levels of factors associated with BRAF, Hippo pathway, and CLDN4. ( F ) Effect of CPE on protein levels of factors associated with EMT and stemness. ( G ) BRAF mutation, nuclear YAP and protein levels of factors associated with EMT were examined in 4 CRC cases with or without CPE expression. Error bar, standard deviation from 3 independent trials. CLDN4 M, membranous CLDN4; CLDN4 C, cytosolic CLDN4; EMT, epithelial-mesenchymal-transition; IC, inhibitory concentration.

Article Snippet: HT29 and HCT116 human OSCC cell lines were purchased from Dainihon Pharmaceutical Co. (Tokyo, Japan).

Techniques: Mutagenesis, Expressing, Standard Deviation, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Malic Enzyme 1 Is Associated with Tumor Budding in Oral Squamous Cell Carcinomas

doi: 10.3390/ijms21197149

Figure Lengend Snippet: Effect of extracellular pH on the epithelial–mesenchymal transition (EMT) phenotype in HSC4 and KON oral squamous cell carcinoma (OSCC) cells. ( A , B ) Effect of the knockdown of ME1 on mRNA expression of EMT-associated genes measured by quantitative PCR ( A ) and extracellular pH ( B ) in HSC4 and KON cells. ( C , D ) Effect of the knockdown of lactate dehydrogenase A (LDHA) on extracellular lactate concentration ( C ) and pH ( D ) in HSC4 and KON cells. ( E ) Effect of medium pH on the EMT phenotype in HSC4 and KON cells. Expression was measured by quantitative PCR. Error bar, standard deviation tested by Student’s t -test in three independent trials. CDH, E-cadherin; CLDN, claudin; HIF, hypoxia-inducible factor; siRNA, small interference RNA; siME, siRNA for ME1; siC, control siRNA; siLDHA, siRNA for LDHA.

Article Snippet: We purchased HSC4 and KON human OSCC cell lines from Dainihon Pharmaceutical Co. (Tokyo, Japan).

Techniques: Knockdown, Expressing, Real-time Polymerase Chain Reaction, Concentration Assay, Standard Deviation, Control

Journal: International Journal of Molecular Sciences

Article Title: Malic Enzyme 1 Is Associated with Tumor Budding in Oral Squamous Cell Carcinomas

doi: 10.3390/ijms21197149

Figure Lengend Snippet: Effect of lactate on the EMT phenotype in HSC4 OSCC cells. ( A – C ) Effect of knockdown of pyruvate dehydrogenase (PDH) or inhibition of PDH kinase by dichloroacetate (DCA) on extracellular lactate ( A ), extracellular pH ( B ), and mRNA expression of EMT-associated genes measured by quantitative PCR ( C ) in HSC4 cells. ( D – F ) Effect of knockdown of monocarbonate transporter (MCT)1 on extracellular lactate ( D ), extracellular pH ( E ), and mRNA expression of EMT-associated genes measured by quantitative PCR ( F ) in HSC4 cells. Error bar, standard deviation tested by Student’s t -test in three independent trials. siPDH, siRNA for PDH; siMCT1, siRNA for MCT1.

Article Snippet: We purchased HSC4 and KON human OSCC cell lines from Dainihon Pharmaceutical Co. (Tokyo, Japan).

Techniques: Knockdown, Inhibition, Expressing, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Malic Enzyme 1 Is Associated with Tumor Budding in Oral Squamous Cell Carcinomas

doi: 10.3390/ijms21197149

Figure Lengend Snippet: Effect of hypoxia on EMT and mitochondrial function in HSC4 and KON OSCC cells. ( A ) Effect of CoCl 2 -induced hypoxia on cell proliferation in HSC4 and KON OSCC cells. ( B ) Effect of hypoxia on mRNA expression of MMPs in HSC4 and KON OSCC cells. ( C ) Effect of hypoxia on mRNA expression of EMT-associated genes. ( D – F ) Effect of hypoxia on mitochondrial membrane potential (TMRE assay). ( D ), mitochondrial reactive oxygen species (ROS, DHR assay) ( E ), and extracellular pH ( F ). Inserts in ( D , E ) show fluorescent images. Error bar, standard deviation tested by Student’s t -test in three independent trials. Scale bar, 50 μm. GAPDH, glyceraldehyde 3-phosphate dehydrogenase; TMRE, tetramethylrhodamine ethyl ester; ROS, reactive oxygen species; DHR123, dihydrorhodamine 123.

Article Snippet: We purchased HSC4 and KON human OSCC cell lines from Dainihon Pharmaceutical Co. (Tokyo, Japan).

Techniques: Expressing, Membrane, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Malic Enzyme 1 Is Associated with Tumor Budding in Oral Squamous Cell Carcinomas

doi: 10.3390/ijms21197149

Figure Lengend Snippet: Effect of ME1 and hypoxia on yes-associated protein (YAP) activation in HSC4 OSCC cells. ( A , B ) Effect of knockdown of ME1 on phosphorylation ( A ) and nuclear translocation ( B ) of YAP in HSC4 cells. ( C ) Effect of CoCl 2 -induced hypoxia on nuclear translocation of YAP in HSC4 cells. ( D ) Concentrations of nuclear YAP were measured by ELISA. Tubulin and lamin were examined as loading controls. pYAP, phosphorylated YAP; LATS, large tumor suppressor; pLATS, phosphorylated LATS; siME, siRNA for ME1.

Article Snippet: We purchased HSC4 and KON human OSCC cell lines from Dainihon Pharmaceutical Co. (Tokyo, Japan).

Techniques: Activation Assay, Knockdown, Phospho-proteomics, Translocation Assay, Enzyme-linked Immunosorbent Assay