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. 2016 Mar 14;212(6):661–676. doi: 10.1083/jcb.201503047

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© 2016 Chen et al.

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Figure 6. — Arl2 functions together with TBCD to regulate microtubule growth and D-TACC/Msps localization. (A) Silver staining of Elution fractions E1 and E2 after TAP. E2 sample contains most of the proteins associated with Arl2-CTAP. Asterisk indicates the bait protein. (B) Coimmunoprecipitation (IP) of Arl2^WT-Venus and FLAG-TBCD, HA-TBCC or HA-TBCE. (C) Coimmunoprecipitation of different forms of Arl2-Venus and HA-TBCC or HA-TBCE. (D) Coimmunoprecipitation of different forms of Arl2-Venus and Myc–β-tubulin or Myc–α-tubulin. (E) NBs of control and TBCD overexpression labeled with α-tubulin and DNA. (F) NBs of control and TBCD overexpression labeled for D-TACC, γ-tubulin, and DNA. Arrows indicate the centrosomes. (G) Metaphase NBs of control, Arl2^T30N, TBCD, and Arl2^T30N, TBCD coexpression labeled with α-tubulin and DNA. (H) Metaphase NBs of control, TBCD overexpression, and TBCD overexpression in heterozygous arl2^Δ156 background (arl2^Δ156/+) labeled for α-tubulin, PH3 and DNA. (E–H) The white dotted circles label the cell outlines. (I) Quantification of spindle length (with SD) in H. Control: 9.48 ± 0.91 µm (n = 16); TBCD: 6.61 ± 1.23 µm (n = 15); TBCD, arl2^Δ156/+: 4.41 ± 0.82 µm (n = 15). ***, P < 0.001. (J) Larval brains of control, TBCD overexpression, and TBCD overexpression in heterozygous arl2^Δ156 background stained with Dpn. Central brain is to the left of the white dotted line. (K) Quantification of NB number per brain hemisphere (with SD) in J. Control: 99 ± 5 (n = 20); TBCD: 149 ± 17 (n = 18); TBCD, arl2^Δ156/+: 369 ± 90 (n = 13). ***, P < 0.001. Bars: (E–H) 5 µm; (J) 20 µm.