anti prl 3  (R&D Systems)

Journal: Biomolecules

Article Title: Credentialing and Pharmacologically Targeting PTP4A3 Phosphatase as a Molecular Target for Ovarian Cancer

doi: 10.3390/biom11070969

Figure Lengend Snippet: PTP4A3 is overexpressed in OvCa cell lines. ( A ) PTP4A3 mRNA expression in the Broad Cancer Cell Library Encyclopedia. Data obtained from https://portals.broadinstitute.org/ccle/ accessed on 31 June 2021. Ovarian cancers are indicated in orange. ( B ) PTP4A3 mRNA levels measured by qPCR and normalized to HPRT were elevated in all HGSOC cells lines (black columns) compared to HIO-180 nonmalignant epithelial cells grown in culture with the exception of COV362 cells. N = 3, bars = SEM. * p < 0.05 ANOVA. demonstrates the differences are not associated with the HPRT reference gene product. ( C ) Detection of PTP4A3 protein in high grade serous OvCa cell lines. Fifty µg of total protein lysate was loaded per lane and protein levels were detected by Western blots with PTP4A3 antibody. Mean values of 2 technical replicates. Bars = range.

Article Snippet: Membranes were blocked in 5% non-fat milk and 2% equine serum in TBS-t for 1 h, and then incubated with the primary PTP4A3 antibody in TBS-t for 1 h followed by secondary HRP-linked anti-mouse antibody in TBS-t for 1 h. Blots were stripped for 3 min using Restore PLUS stripping buffer, rinsed with TBS-t, blocked with 5% non-fat milk in TBS-t for 1 h, then incubated with loading control α-tubulin antibody in TBS-t for 1 h followed by secondary HRP-linked anti-antibody in TBS-t for 1 h. We used the following commercially available primary and secondary antibodies: PTP4A3 (Novus Biologicals, Centennial, CO, USA, catalog # MAB3219), anti-mouse IgG HRP-linked (Cell Signaling Technology, Danvers, MA, USA, catalog #7076) α-tubulin (Cell Signaling Technology, catalog #2125), anti-rabbit IgG HRP-linked (Cell Signaling Technology, catalog #7074).

Techniques: Expressing, Western Blot

Journal: Biomolecules

Article Title: Credentialing and Pharmacologically Targeting PTP4A3 Phosphatase as a Molecular Target for Ovarian Cancer

doi: 10.3390/biom11070969

Figure Lengend Snippet: PTP4A3 overexpression in human OvCa is associated with poor survival, even in early stage disease. ( A ) High PTP4A3 expression was associated with poor survival in tumors from 98 patients with stage I/II serous OvCa. Red symbols and line represents the 48 tumor samples with high PTP4A3 expression and black symbols and line represent the 50 tumor samples with low PTP4A3 expression. Patients with low PTP4A3 gene expression showed almost double the length of overall survival: 81.2 vs. 44.3 months. p < 0.05. ( B ) Beeswarm plot showing high expression of PTP4A3 even in early stage OvCa patients. Red and black symbols are from Panel A. ( C ) High PTP4A3 expression is also associated with reduced survival in stage III/IV serous OvCa patents. Red symbols and line represents the 394 tumor samples with high PTP4A3 expression and black symbols and line represent the 629 tumor samples with low PTP4A3 expression. Patients with low PTP4A3 expression had an overall survival of 44.5 months compared to 36.8 months for patients with high PTP4A3 expressing tumors. p < 0.05. ( D ) Beeswarm plot showing high expression of PTP4A3 maintained in a significant population of patients in later stage disease. Red and black symbols are from Panel ( C ).

Article Snippet: Membranes were blocked in 5% non-fat milk and 2% equine serum in TBS-t for 1 h, and then incubated with the primary PTP4A3 antibody in TBS-t for 1 h followed by secondary HRP-linked anti-mouse antibody in TBS-t for 1 h. Blots were stripped for 3 min using Restore PLUS stripping buffer, rinsed with TBS-t, blocked with 5% non-fat milk in TBS-t for 1 h, then incubated with loading control α-tubulin antibody in TBS-t for 1 h followed by secondary HRP-linked anti-antibody in TBS-t for 1 h. We used the following commercially available primary and secondary antibodies: PTP4A3 (Novus Biologicals, Centennial, CO, USA, catalog # MAB3219), anti-mouse IgG HRP-linked (Cell Signaling Technology, Danvers, MA, USA, catalog #7076) α-tubulin (Cell Signaling Technology, catalog #2125), anti-rabbit IgG HRP-linked (Cell Signaling Technology, catalog #7074).

Techniques: Over Expression, Expressing, Gene Expression

Journal: Biomolecules

Article Title: Credentialing and Pharmacologically Targeting PTP4A3 Phosphatase as a Molecular Target for Ovarian Cancer

doi: 10.3390/biom11070969

Figure Lengend Snippet: Chemical structures of the inhibitors and in vitro IC 50 against recombinant human PTP4A3. Mean ± SEM, N = 3, unless indicated by * N = 6 or ** N = 8.

Article Snippet: Membranes were blocked in 5% non-fat milk and 2% equine serum in TBS-t for 1 h, and then incubated with the primary PTP4A3 antibody in TBS-t for 1 h followed by secondary HRP-linked anti-mouse antibody in TBS-t for 1 h. Blots were stripped for 3 min using Restore PLUS stripping buffer, rinsed with TBS-t, blocked with 5% non-fat milk in TBS-t for 1 h, then incubated with loading control α-tubulin antibody in TBS-t for 1 h followed by secondary HRP-linked anti-antibody in TBS-t for 1 h. We used the following commercially available primary and secondary antibodies: PTP4A3 (Novus Biologicals, Centennial, CO, USA, catalog # MAB3219), anti-mouse IgG HRP-linked (Cell Signaling Technology, Danvers, MA, USA, catalog #7076) α-tubulin (Cell Signaling Technology, catalog #2125), anti-rabbit IgG HRP-linked (Cell Signaling Technology, catalog #7074).

Techniques: In Vitro, Recombinant

Journal: Biomolecules

Article Title: Credentialing and Pharmacologically Targeting PTP4A3 Phosphatase as a Molecular Target for Ovarian Cancer

doi: 10.3390/biom11070969

Figure Lengend Snippet: Inhibition of PTP4A3 activity blocks OvCa cell migration. ( A ) JMS-053 inhibits migration of A2780 cells. A2780 were cultured to confluence then “wounded” and treated with vehicle control or JMS-053. Wound closure was quantified at 14 h post-wounding. Mean ± SEM ( B ) IL-6 caused a concentration-dependent migration in PTP4A3 wildtype OVCAR4 cells (blue). CRISPR/Cas9 knock-out of PTP4A3 in OVCAR4 cells reduces IL-6 mediated cell migration (red). Mean ± SEM. ( C ) JMS-053 causes a concentration-dependent inhibition of IL-6-mediated migration in wildtype OVCAR4 cells. Wound closure was quantified at 18 h post-wounding. Mean ± SEM. N = 3. Vehicle control indicated by the red line. ( D ). With OVCAR4-PTP4A3 knockout cells, JMS-053 did not cause any further inhibition of migration in the presence of IL-6. Wound closure was quantified at 18 h post-wounding. Mean ± SEM, N = 3. * p < 0.05.

Article Snippet: Membranes were blocked in 5% non-fat milk and 2% equine serum in TBS-t for 1 h, and then incubated with the primary PTP4A3 antibody in TBS-t for 1 h followed by secondary HRP-linked anti-mouse antibody in TBS-t for 1 h. Blots were stripped for 3 min using Restore PLUS stripping buffer, rinsed with TBS-t, blocked with 5% non-fat milk in TBS-t for 1 h, then incubated with loading control α-tubulin antibody in TBS-t for 1 h followed by secondary HRP-linked anti-antibody in TBS-t for 1 h. We used the following commercially available primary and secondary antibodies: PTP4A3 (Novus Biologicals, Centennial, CO, USA, catalog # MAB3219), anti-mouse IgG HRP-linked (Cell Signaling Technology, Danvers, MA, USA, catalog #7076) α-tubulin (Cell Signaling Technology, catalog #2125), anti-rabbit IgG HRP-linked (Cell Signaling Technology, catalog #7074).

Techniques: Inhibition, Activity Assay, Migration, Cell Culture, Control, Concentration Assay, CRISPR, Knock-Out