Cell division

A549, H460, H1650 are non-small cell lung cancer lines

1. % mitotic cells compared to control through siRNA transfection

2. Knockdown using lentiviral shRNA

3. Western blot; cells synchronized at G1/S, released for varying times

   1. p-TBK1 activated 4 hours post release

   2. Maximal TBK1 phosphorylation= maximal Histone H3 phosphorylation

D) BX795 inhibits TBK1; control cells at 9 hours post release had pH3S10, but treated cells didn’t

E) TBK1 depleted by siRNA transfection, no pH3S10 (small band at 9hr, not mentioned in paper)

F) PLK1 is mitotic kinase, may be substrate of TBK1;Testing if overexpressing PLK1 can overpower depleted TBK1, but it can’t rescue from TBK1 depletion; TBK1 must target other proteins to modulate mitosis