michael pacold  (Addgene inc)

Journal: Cancer discovery

Article Title: Limited Environmental Serine and Glycine Confer Brain Metastasis Sensitivity to PHGDH Inhibition

doi: 10.1158/2159-8290.CD-19-1228

Figure Lengend Snippet: (A) Radiance of brain metastasis-bearing mice measured at 28 days following intracardiac injection of aggressive TNBC BrM cells expressing either a control shRNA hairpin or shRNA hairpin targeting PHGDH. (B) Kaplan-Meier plot showing disease-specific survival of mice injected with Aggressive TNBC BrM cells expressing either a control shRNA (shCtrl) or PHGDH shRNA (shPHGDH #1 or shPHGDH #2). Significance was tested using the log-rank test. (C) Radiance of brain metastasis-bearing mice measured at 28 days following intracardiac injection of NSCLC aggressive BrM cells expressing either a control shRNA hairpin or shRNA hairpin suppressing PHGDH. (D) Kaplan-Meier plot showing disease-specific survival of mice injected with NSCLC aggressive BrM cells expressing either a control shRNA (shCtrl) or PHGDH shRNA (combined shPHGDH #1 and shPHGDH #2). Significance was tested using the log-rank test. (E) Representative images of GFP-positive brain metastatic lesions derived from tumor bearing mice injected with parental MDA-MB-231 cells expressing either an empty vector (EV) control, catalytically active PHGDH, or catalytically inactive PHGDH. (F) Quantitation of brain metastases from parental MDA-MB-231 brain metastases by direct GFP visualization of isolated brains. (G) Radiance of mice measured at 28 days following intracardiac injection with parental MDA-MB-231 cells expressing an empty vector control, catalytically active PHGDH or catalytically inactive PHGDH.

Article Snippet: Catalytically inactive PHGDH constructs are available from Addgene as follows: pCW-codon optimized catalytically inactive PHGDH (154903) and pMXS-catalytically inactive PHGDH (154906).

Techniques: Injection, Expressing, Control, shRNA, Derivative Assay, Plasmid Preparation, Quantitation Assay, Isolation

Journal: Cancer discovery

Article Title: Limited Environmental Serine and Glycine Confer Brain Metastasis Sensitivity to PHGDH Inhibition

doi: 10.1158/2159-8290.CD-19-1228

Figure Lengend Snippet: (A) Fractional labeling of intracellular U-13C-glucose-derived m+3 serine from aggressive or indolent brain metastatic cells grown in Plasma, CSF, or -Ser/-Gly media. (B) Proliferative capacity of aggressive or indolent TNBC BrM cells grown in Plasma, CSF, or -Ser/-Gly media. (C) Fractional labeling of intracellular U-13C-glucose-derived serine from aggressive brain-trophic cell lines expressing a control shRNA (shCtrl) or PHGDH shRNA (shPHGDH #1 or shPHGDH #2) (D) Proliferative capacity of aggressive TNBC BrM cells expressing a control shRNA (shCtrl) or PHGDH shRNA (shPHGDH #1 or shPHGDH #2). (E) Fractional labeling of intracellular U-13C-glucose -derived serine in aggressive TNBC BrM cells with a PHGDH shRNA (shPHGDH #2) expressing either an empty vector (EV) control, catalytically active or catalytically inactive, shRNA-resistant PHGDH. (F) Proliferative capacity of PHGDH knockdown aggressive TNBC BrM cells with a PHGDH shRNA (shPHGDH #2) expressing either an empty vector (EV) control, catalytically active or catalytically inactive, shRNA-resistant PHGDH. Cell proliferation data was monitored over 4-6 days using an Incucyte or Multisizer Coulter Counter. Error bars represent standard deviations.

Article Snippet: Catalytically inactive PHGDH constructs are available from Addgene as follows: pCW-codon optimized catalytically inactive PHGDH (154903) and pMXS-catalytically inactive PHGDH (154906).

Techniques: Labeling, Derivative Assay, Clinical Proteomics, Expressing, Control, shRNA, Plasmid Preparation, Knockdown