Fig. 2.

Mutant PRKCA induces ERK phosphorylation and is sufficient to drive anchorage-independent growth of immortalized human astrocytes. a Colony formation in soft agar of immortalized human astrocytes after infection with lentivirus expressing empty vector, wildtype PRKCA, D463H tumor mutant PRKCA, or D463A non-tumor mutant PRKCA. Images of representative wells (left) and quantitation of colony number per well (right) are shown. Error bars represent standard deviation from the mean of six replicates derived from two independent experiments performed in triplicate. b Western blots on total cell lysate from 293 T cells growing in 10% fetal bovine serum after transient transfection with empty vector, PRKCA wild-type, or PRKCA D463H mutant. c Western blots on immortalized human astrocytes after lentiviral transduction and then serum starvation for 24 h before collection of total cell lysate