Cryo-CLEM analysis of kinesore-induced projections. HAP1 cells grown on Quantifoil gold EM grids were stained with Cell Mask green to identify plasma membrane and treated with kinesore (100 µM) for 1 h before plunge-freezing and imaging using fluorescence and EM. (A) Low magnification images show fluorescence (green) and EM image (grays) overlay of cells with projections emerging. Regions boxed i and ii are shown at higher magnification and expanded on the right. Black dots are 10 nm gold fiducials. (B) A single EM image acquired proximal to the cell body with a large number of microtubules. Boxes show microtubules with similar alignments (shaded blue, 1–3 [top] and 1–4 [bottom]) and an example of a single microtubule that crosses a bundle of aligned microtubules (yellow, 5 [bottom]). Scale bars are 100 nm.