Characterisation of linear, hPOC5-induced structures.

<div><p>Immunofluorescence micrographs show examples of the structures induced by stable expression of hPOC5-GFP in U2OS cells. GFP is shown in the green channel, with relevant markers in red and DNA visualised with DAPI (blue). Blow-ups (2.5x) show the hPOC5-induced structures. Micrographs are representative of results obtained in at least 3 separate experiments. Scale bar, 5 µm.</p> <p>A. Centrin2 and centrin3 (red) localise at the hPOC5-induced structures.</p> <p>B. hPOC5-induced structures are observed in a proportion of mitotic cells but do not contain α-tubulin.</p> <p>C. hPOC5-induced structures contain γ-tubulin and NEDD1, but not acetylated tubulin (stabilized microtubules).</p> <p>D. PCM1, but not pericentrin, is seen at hPOC5-induced structures.</p> <p>E. hPOC5 structures are resistant to microtubule depolymerisation. Cells were treated with 2µg/ml nocodazole for 2h before fixation.</p> <p>F. Electron micrographs of (<b>i, ii, iv, v</b>) U2OS cells with stable expression of hPOC5 showing (<b>i</b>) normal cross-sectional centriole structure; examples of elongated centriolar microtubules in (ii) U2OS and (iii) DT40 cells that overexpressed POC5; (<b>iv</b>)<b>, (v)</b> examples of electron-dense, linear aggregates (<b>vi</b>). Micrograph shows a primary cilium in hTERT-RPE1 cells. Scale bar, 200 nm.</p>