Figure 1.
Figure 1. SFs promote BB reorientation. (A) The BB reorientation assay. BB disorientation was exacerbated by shifting DISA-1 rescue cells from 25°C to 37°C for 24 h. To promote BB reorientation, cells were shifted from 37°C to 25°C coincident with WT DisAp protein expression. (B) Schematic of SF length measurements, distance measurements from the posterior BB to the anterior BB’s pcMT distal end to establish the minimal SF length that is required for SF–pcMT contact, and BB orientation analyses. BBs and pcMTs, red; SF, green. (C) i and ii: DisAp expression leads to SF elongation before BB reorientation. BB reorientation occurs when SF length surpasses the minimal length that is required for SF–pcMT interactions (arrowhead and dotted line marks the mean distance from the posterior BB to the anterior BB’s pcMT distal end in WT cells). SF length and BB orientation partially recover 2 wk after DISA-1 rescue. BB, red; SF, green. All small insets show a representative BB and SF. i, bottom: White box marks region of interest (large inset). iii, top: Average SF length positively corresponds with BB orientation (R value) on the single cell level during DISA-1 rescue. iii, bottom: Correlation analysis between the average SF length and BB orientation during DISA-1 rescue. BB orientation in most cells is recovered once SF length surpasses the minimal length that is required to establish SF–pcMT contacts (dotted line marks the mean distance from the posterior BB to the anterior BB’s pcMT distal end in WT cells). Data points are fitted with a polynomial (order = 2) function. n ≥ 300 SFs (≥30 cells). Mann-Whitney test. * denotes P value <0.01. Mean ± SD. Scale bars, 10 µm (cell), 1.3 µm (small inset width), 7.8 µm (large inset width). (D) Frequency of SF–pcMT contacts is increased as SF length recovers in DISA-1 rescue cells. Schematic illustrates the position and orientation of two BBs within a region of interest (white box). BB-associated microtubule appendages, red; SF, green. n = 240 SFs (24 cells). Scale bar, 5 µm. tMT, transverse microtubule.

SFs promote BB reorientation. (A) The BB reorientation assay. BB disorientation was exacerbated by shifting DISA-1 rescue cells from 25°C to 37°C for 24 h. To promote BB reorientation, cells were shifted from 37°C to 25°C coincident with WT DisAp protein expression. (B) Schematic of SF length measurements, distance measurements from the posterior BB to the anterior BB’s pcMT distal end to establish the minimal SF length that is required for SF–pcMT contact, and BB orientation analyses. BBs and pcMTs, red; SF, green. (C) i and ii: DisAp expression leads to SF elongation before BB reorientation. BB reorientation occurs when SF length surpasses the minimal length that is required for SF–pcMT interactions (arrowhead and dotted line marks the mean distance from the posterior BB to the anterior BB’s pcMT distal end in WT cells). SF length and BB orientation partially recover 2 wk after DISA-1 rescue. BB, red; SF, green. All small insets show a representative BB and SF. i, bottom: White box marks region of interest (large inset). iii, top: Average SF length positively corresponds with BB orientation (R value) on the single cell level during DISA-1 rescue. iii, bottom: Correlation analysis between the average SF length and BB orientation during DISA-1 rescue. BB orientation in most cells is recovered once SF length surpasses the minimal length that is required to establish SF–pcMT contacts (dotted line marks the mean distance from the posterior BB to the anterior BB’s pcMT distal end in WT cells). Data points are fitted with a polynomial (order = 2) function. n ≥ 300 SFs (≥30 cells). Mann-Whitney test. * denotes P value <0.01. Mean ± SD. Scale bars, 10 µm (cell), 1.3 µm (small inset width), 7.8 µm (large inset width). (D) Frequency of SF–pcMT contacts is increased as SF length recovers in DISA-1 rescue cells. Schematic illustrates the position and orientation of two BBs within a region of interest (white box). BB-associated microtubule appendages, red; SF, green. n = 240 SFs (24 cells). Scale bar, 5 µm. tMT, transverse microtubule.

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