Journal: Cancers

Article Title: Discovery of Functional Alternatively Spliced PKM Transcripts in Human Cancers

doi: 10.3390/cancers13020348

Figure Lengend Snippet: Biological effect of PKM transcript over-expression in PC3 human prostate cancer cell line. ( a ) Western blots image showing the over-expression of transfected PKM-093 and PKM-883, respectively. ( b ) Pyruvate kinase activity kinetic assay. Kinetic PK activity was measured using 7.5 µg of each transfected cell lysate. PKM-093 and PKM-883 reached faster plateau status at 6 and 8 min, respectively, than pCMV6KN empty vector control which reached plateau at 10 min. ( c ) Western blots for the overexpressed proteins of PKM-093 and PKM-883 in cytosol and nucleus with the same protein volume (see Method ). The red dot means cytosolic protein and the green dot means nuclear protein. ( d ) Bar chart and microscopy images showing the over-expression of PKM-093 and PKM-883 significantly inhibited cell invasion compared to negative control. Error bars indicate standard errors. * indicates t -test p < 0.05. Scale bar = 100 micrometer (µm). ( e ) Quantitative PCR analysis showing the mRNA level changes of SLC2A1, PDK1, and SREBF1 in PC3 cells with PKM-093 and PKM-883 over-expression compared to negative control. Error bars indicate standard errors. * indicates t -test p < 0.05.

Article Snippet: SDS PAGE separated 60 μg of PC3 cell lysate per well with Precision Plus Protein Standards ladder (1610374, Bio-Rad).

Techniques: Over Expression, Western Blot, Transfection, Activity Assay, Kinetic Assay, Plasmid Preparation, Control, Microscopy, Negative Control, Real-time Polymerase Chain Reaction