Light chain 2 is a Tctex-type related axonemal dynein light chain that regulates directional ciliary motility in Trypanosoma brucei
Fig 2
Overexpressed TbLC2 localized to the axoneme as well as the cytoplasm.
A. Doxycycline induced TbLC2::eGFP overexpressing wild-type cells (WT/LC2 OE cell) visualized using differential interference contrast (DIC) microscopy (left) and under widefield fluorescence microscopy (right). The TbLC2::eGFP (green) localized to the cytoplasm and the cilium. The immunostained paraflagellar rod 2 (PFR2, red), DAPI stained DNA (DAPI, blue), and TbLC2::eGFP overlay (right), suggest that the TbLC2 has ciliary and cytoplasmic localization. B. SEM images showing cilium and cell body morphology of wild-type (WT, left), FLAM3-LC2 KD (middle), and FLAM3-LC2 KD/LC2 OE (right) cells showing short flagellar attachment zones (arrows) and nearly complete ciliary detachment from the cell body in the FLAM3 knocked down cells. The scale bar is 5 μm in each panel. C. LC2 KD (left), FLAM3 KD (center), and FLAM3-LC2 KD (right) cells visualized with DIC microscopy showing short flagellar attachment zones (arrows) and nearly complete ciliary detachment from the cell body. D. TbLC2::eGFP (green) expressing FLAM3-LC2 KD/LC2 OE cells stained with anti-ɑ-tubulin antibody (red) visualized with a confocal microscope. E. TbLC2::eGFP (green) expressing FLAM3-LC2 KD/LC2 OE cells stained with DAPI (blue) and the anti-PFR2 antibody (red) visualized under a widefield fluorescence microscope. The inset shows that TbLC2::eGFP and the paraflagellar rod are separately localized, and the line scan plots shows the normalized fluorescence intensity of TbLC2::eGFP (green) and PFR2 (red) along the line (white) shown in the inset.