The Pea2-Myo2 complex can walk on actin cables. (A) Left: Experimental setup. Pea2 was fused to a fragment of Pex3 and thus coupled to peroxisomes (Pex3_1–35-mCherry-Pea2 [Pex-Pea2]). Right: WT cells or myo2_RD cells coexpressing Pex or Pex-Pea2 together with the peroxisomal import substrate GFP-SKL were analyzed by two-channel fluorescence microscopy. Scale bar: 5 µm. (B) mCherry-stained peroxisomes were quantified in WT or myo2_RD cells expressing either Pex (n_WT = 228; n_myo2RD = 215), or Pex-Pea2 (n_WT = 214; n_myo2RD = 215). ns, not significant; ***, P < 0.001 (one-way ANOVA followed by Dunn’s multiple comparison test). (C) The distribution of mCherry-stained peroxisomes between mother and daughter cells was determined in WT or myo2_RD cells expressing either Pex (n_WT = 228; n_myo2RD = 215) or Pex-Pea2 (n_WT = 214; n_myo2RD = 215).