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. 2016 Feb 1;27(3):549–571. doi: 10.1091/mbc.E15-11-0754

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© 2016 Liu, He, et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 10: — The coating proteins protect the stability of cortical microtubules as an ensemble. (A) Deconvolved wide-field images of cold-treated parental RHΔhx parasites and various coating protein–knockout mutants labeled with a rabbit anti–Tgβ-tubulin antibody (Morrissette and Sibley, 2002b). Intracellular parasites were incubated at 8°C for 3.5 h before processing for immunofluorescence. In the Δtlap2Δspm1Δtlap3-TLAP3 rescue parasites, mNeonGreenFP-TLAP3 expression was driven by the 2-kb genomic region immediately upstream of tlap3 in the plasmid pTKO2_II_mNeonGreenFP-TLAP3. Insets, 2×. Arrows indicate residual cortical labeling by the anti–Tgβ-tubulin antibody in the Δtlap2Δspm1, Δtlap2Δspm1Δtlap3, or Δtlap2Δspm1Δtlap3- TLAP3 rescue parasites. (B, C) Deconvolved wide-field images of intracellular parental RHΔhx, Δtlap2Δspm1, and Δtlap2Δspm1Δtlap3 parasites labeled with a rabbit anti–Tgβ-tubulin antibody. The parasites were cultured under standard culture conditions (i.e., with no cold treatment) before processing for immunofluorescence. (B) Interphase parasites. (C) Parasites with forming daughters (arrows, mitotic spindle; green, anti-ISP1; red, anti–Tgβ-tubulin).