Mitochondria detach from the cell cortex during meiosis II. Video montages and quantifications of cells expressing Cit1-GFP or Cit1-mCardinal to label mitochondria (mito), as well as a meiotic staging marker, imaged every 10 min. Mitochondrial detachment is defined as the abrupt coalescence of mitochondria, showing restricted rather than uniform localization around the cell cortex (arrowhead). Dashed lines: cell boundaries. To determine the relative staging compared with markers of meiotic progression (below), mitochondrial detachment is defined to occur at 0 min. Plots show the mean ± range (shaded region) of two independent experiments (n ≥ 90 cells counted per experiment per marker). (A) Mitochondrial detachment relative to the onset of the meiosis II nuclear division (anaphase II), marked by Htb1-mCherry (UB10257). Anaphase II is defined as the first appearance of a four-lobed nuclear morphology (*). (B) Mitochondrial detachment relative to prospore membrane nucleation and closure, marked by the GFP-Spo20^51–91 prospore membrane marker (UB13131). Prospore membrane nucleation is defined as the first appearance of Spo20^51–91 puncta (*) and closure as the rounding up of fully elongated prospore membranes (◊). (C) Mitochondrial detachment relative to metaphase II and anaphase II, marked by Spc42-GFP (UB13129). Metaphase II is defined as the first appearance of two pairs of separated Spc42-GFP dots (*). Anaphase II is defined as the first appearance of concerted movement separating the sister spindle pole bodies in each pair (◊). Scale bars, 2 µm.