Journal: Journal of Clinical Medicine

Article Title: Epithelial-Mesenchymal Transition-Related MicroRNAs and Their Target Genes in Colorectal Cancerogenesis

doi: 10.3390/jcm8101603

Figure Lengend Snippet: Probes used for miRNAs and mRNAs quantification using quantitative real-time PCR (qPCR).

Article Snippet: PTPN13 , Hs01106214_m1 , 65.

Techniques: Real-time Polymerase Chain Reaction

Journal: Journal of Clinical Medicine

Article Title: Epithelial-Mesenchymal Transition-Related MicroRNAs and Their Target Genes in Colorectal Cancerogenesis

doi: 10.3390/jcm8101603

Figure Lengend Snippet: Statistically significant correlations between expression of miR-200 family and their target genes and association with E-cadherin staining.

Article Snippet: PTPN13 , Hs01106214_m1 , 65.

Techniques: Expressing, Staining

Journal: Biomedicines

Article Title: Reversal of Myofibroblast Apoptosis Resistance and Collagen Deposition by Phaseoloidin-Induced Autophagy Attenuates Pulmonary Fibrosis

doi: 10.3390/biomedicines13112679

Figure Lengend Snippet: PTPN13 mediated by phaseoloidin links autophagy and apoptosis. ( A ) Western blot analysis of PTPN13 levels in FasL and different dose phaseoloidin co-treated myofibroblasts ( left ) and quantification of the PTPN13 band intensity ( right ). ( B ) Western blotting reflects the CO-IP assay of PTPN13 binding to p62 under co-treatment with FasL and phaseoloidin or CQ. ( C ) Western blot analysis of PTPN13 levels in FasL and 100 μM phaseoloidin co-treated primary mouse myofibroblasts in the presence or absence of CQ ( top ) and quantification of the PTPN13 band intensity ( bottom ). ( D ) Flow cytometry was used to detect the percentage of apoptosis in the above experimental groups. ( E ) Flow cytometry was used to detect the percentage of apoptosis in groups with or without vector-PTPN13. Data in this figure represent the means ± S.D. (* p < 0.05, ** p < 0.01, **** p < 0.0001).

Article Snippet: A total of 10% of the proteins electrophoresed on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels were transferred to PVDF membranes (Bio-Rad) and 5% ( w / v ) skim milk with TBS buffer containing 0.1% ( v / v ) Tween 20 (TBST) TBS buffer for blocking. β-ACTIN monoclonal antibody (1:5000, Proteintech, Wuhan, China), α-SMA monoclonal antibody (1:1000, CST, USA), Caspase3 polyclonal antibody (1:1000, Proteintech, China), collagen I polyclonal antibody (1:1000, Millipore, USA), Bcl-2 polyclonal antibody (1:1000, Proteintech, China), BAX polyclonal antibody (1:1000, Proteintech, China), PTPN13 polyclonal antibody (1:1000, RD, Minneapolis, MN, USA), p70S6K polyclonal antibody (1:1000, CST, USA), elf-4B polyclonal antibody (1:500, ABclonal, Wuhan, China), mTOR monoclonal antibody (1:1000, CST, USA), Pho-mTOR monoclonal antibody (1:1000, CST, USA), AMPK monoclonal antibody (1:1000, CST, USA) and Pho-AMPK monoclonal antibody (1:1000, CST, USA) were incubated overnight at 4 °C.

Techniques: Western Blot, Co-Immunoprecipitation Assay, Binding Assay, Flow Cytometry, Plasmid Preparation

Journal: PLoS ONE

Article Title: Pair-Wise Regulation of Convergence and Extension Cell Movements by Four Phosphatases via RhoA

doi: 10.1371/journal.pone.0035913

Figure Lengend Snippet: (a) Protein structures are shown encoded by ptpn20 homologue and the immediately 5′ upstream FRMPD2 , as currently annotated in five fish genomes, the human genome and the mouse genome. In some cases like Fugu and Tetraodon a single known coding transcript exists besides separate transcripts encoding the PTP domain and the “FRMPD" part. For comparison the protein structure encoded by human ptpn13 (PTPBL) is added below. (b) Primers were designed as indicated, leaving approximately 100 bp known coding sequence for the purpose of alignment of generated sequences. PCR products with forward primers on the second to last known exon of human and zebrafish FRMPD2 and reverse oligos on the second exon of PTPN20 . A schematic representation of retrieved sequences blasted to the genome are indicated in green (not to scale). (c) Generated PCR products on human (top) and zebrafish (bottom) cDNA libraries using the described primer sets. Generated band sizes are consistent with expected values based on homology with the ptpn13 gene.

Article Snippet: Ptpn13 RNA was transcribed from full length mouse cDNA kindly provided by Wiljan Hendriks (Department of Cell Biology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands).

Techniques: Comparison, Sequencing, Generated

Journal: PLoS ONE

Article Title: Pair-Wise Regulation of Convergence and Extension Cell Movements by Four Phosphatases via RhoA

doi: 10.1371/journal.pone.0035913

Figure Lengend Snippet: Morpholinos targeting ptpn13 and ptpn20 were injected in the zebrafish at the one cell stage, and concentrations were titrated down until no phenotype was observed. Normal (red), low (green) concentrations and combined low concentrations of ptpn13 and ptpn20 morpholino were micro-injected and embryos were grown to 3dpf under normal conditions. Pictures were taken from all embryos and tails were measured using ImageJ imaging software, from the yolk to the tip of the tail, and compared to non-injected control. Average tail length compared to non-injected control is plotted as a percentage deviating from 100% in (a) and representative fish are shown for each condition in (b). Zebrafish embryos were microinjected as described above, using low concentration combined knockdown of ptpra with either ptpn13 or ptpn20 , or ptpre with either ptpn13 or ptpn20 and tail lengths are plotted in (c) and (d). (e) Shown are representative fish from the experiments depicted in (c) and (d). All error bars are standard error of the mean. Student t-test was performed where indicated; no asterisk indicates P>0.05, * indicates 0.05>P>0.001 and ** indicates P<0.001. Morpholino concentrations are color coded: red for “full" knockdown, giving full phenotype without being toxic and green for “low" concentration, giving no observable phenotype.

Article Snippet: Ptpn13 RNA was transcribed from full length mouse cDNA kindly provided by Wiljan Hendriks (Department of Cell Biology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands).

Techniques: Injection, Imaging, Software, Control, Concentration Assay, Knockdown

Journal: PLoS ONE

Article Title: Pair-Wise Regulation of Convergence and Extension Cell Movements by Four Phosphatases via RhoA

doi: 10.1371/journal.pone.0035913

Figure Lengend Snippet: (a) Low dose combined knockdowns of ptpn13 or ptpn20 and arhgap29b were performed by injecting indicated amounts of morpholino at the one cell stage. Tail lengths were measured at 3dpf and plotted. Co-knockdowns with arhgap5 were included as a control. (b) Similar co-knockdowns as in (a) but with ptpra and ptpre knockdown instead of ptpn13 and ptpn20 knockdown. (c) Zebrafish embryos were micro-injected with morpholinos targeting the different phosphatases in low concentrations together with low dose arhgef27 ( ngef ) morpholino. Embryos were grown to 3 dpf and tail lengths were determined and plotted as a percentage of non-injected control. All error bars are standard error of the mean. Student t-test was performed where indicated; no asterisk indicates P>0.05, * indicates 0.05>P>0.001 and ** indicates P<0.001.

Article Snippet: Ptpn13 RNA was transcribed from full length mouse cDNA kindly provided by Wiljan Hendriks (Department of Cell Biology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands).

Techniques: Control, Knockdown, Injection