Effects of shRNA targeting nestin on cell morphology. (A) Sham-transfected PANC-1 and PK-45H cells were either round or had a short spindle-like appearance. Nestin shRNA-transfected cells exhibited a sheet-like appearance with the cells clustering to each other. (B) There was also increased F-actin expression at the periphery of the cells, and a dramatic increase in stress fiber formation of F-actin by comparison with sham cells. (C) expression patterns of α-tubulin were similar in sham and shRNA-transfected cells. (D) Merged images of F-actin and α-tubulin showed strong co-expression of the proteins (yellow) at the periphery of nestin shRNA-transfected PANC-1 and PK-45H cells. (A) phase-contrast images of live cells (original magnification, ×200); (B) immunofluorescence of F-actin (red, original magnification, ×1,000); (C) immunofluorescence of α-tubulin (green, original magnification, ×1,000); (D) Merged images of F-actin and α-tubulin (yellow, Original magnification, ×1,000).