Supplemental Materials

This article contains the following supporting material:

• Supplemental Figure 1 - No colocalization of PAK5 with cell organelles:

  CHO cells transfected with YFP-PAK5 wt were cultured for 16 h, fixed and costained with different markers. No colocalization was found with markers of lysosomes (LysoTracker), peroxisomes (antibody against PMP69), and mitochondria (MitoTracker).

• Supplemental Figure 2 - No colocalization of PAK5/MARK2-vesicles and vimentin filaments:

  CHO cells transfected with YFP-PAK5^MM (yellow) and CFP-MARK2^E (cyan) constructs were cultured for 16 h, fixed and costained with anti-vimentin- and Cy5-secondary antibody (red). Immunostaining of vimentin filaments shows no vimentin cage-like structures in cells transfected with inactive PAK5^MM and active MARK2^E.

• Supplemental Figure 3 - Effects of PAK5 and MARK2 on the stability of microtubules:

  CHO cells transfected with active YFP-PAK5^NE were cultured for 16 h, fixed and costained with Glu-tubulin-antibody and Cy5-secondary antibody (MT (Glu)-staining, red). Transfected cells are indicated by arrows. Constitutively active PAK5 (1, green) stabilizes MT (glu) (2, red).

• Supplemental Figure 4 - Effects of PAK5-AID on MARK2 and the stability of microtubules:

  CHO cells transfected with myc-tagged PAK5-AID (unable to bind MARK2, stained with Cy5-secondary antibody) alone or together with CFP-MARK2 were cultured for 16 h, fixed and costained with YL1/2- and TRITC-secondary antibody (MT-staining, red). Transfected cells are indicated by arrows. Cells expressing PAK5-AID alone (1, green) show a normal microtubule network (2). In cells coexpressing PAK5-AID (4, yellow) and wildtype CFP-MARK2 (5, cyan), microtubules dissappear (6, arrow). The AID-domain of PAK5 is unable to block MARK2.

• Supplemental Figure 5 - PAK5 inhibits MARK2 induced tau phosphorylation:

  CHO-cells stably transfected with htau40 were transiently transfected with YFP-PAK5^MM and CFP-MARK2, cultured for 16 h, fixed and costained with phospho-tau antibody (12E8) (Cy5-secondary antibody, red). Transfected cells are indicated by arrows. Expression of MARK2 wt (1, cyan) results in a strong Tau phosphorylation (2, red). The inactive form of PAK5 (4, yellow) shows no Tau phosphorylation. Coexpression of inactive PAK5 (7, yellow) and active MARK2 (8, cyan) results in an inhibition of MARK2. The Tau phosphorylation is strongly reduced (9, red).

• Supplemental Material