Localizations and behaviors of exogenous Kif6 or Kif9 in ependymal cilia. Related to Figs. 2 and 3. (A) Additional examples for motility behaviors of ciliary Kif6-GFP and Kif9-GFP. mEPCs were infected with adenovirus as illustrated in Fig. 2 A to express the indicated GFP-tagged proteins. Ift81-GFP served as a positive control. GI-SIM images were recorded and processed as described in Fig. 2, C–E. Arrows indicate typical clearly traceable GFP-positive puncta to show their movements. (B) 3D-SIM images showing spatial relationships between Kif6-GFP and Ift56 or Ift88. Acetylated tubulin (Ac-tub) marked ciliary axonemes. The cilia magnified from the framed regions are presented in Fig. 3 A. (C) Typical images of microscope calibrations. Instrument calibration was conducted routinely with fluorescent beads prior to dual-color GI-SIM imaging experiments. A line scan was performed along the white line to determine the alignment. (D) Additional examples for bidirectional movements of ciliary Kif6-GFP-alone (Kif6⁺) and Ift27-2Halo-alone (Ift27⁺) puncta. mEPCs were treated and imaged as described in Fig. 3, B–F.