Figure 1.
Figure 1. Kinesin-1 and cytoplasmic dynein function in an interdependent manner during peroxisome transport in S2 cells. (A) Potential outcomes of RNAi-based motor depletion if motors work independently of each other. Schematic depicts S2 cells plated in the presence of 1 µM CytoD to induce formation of microtubule-filled processes. Microtubule polarity is indicated with + and − signs. Green dots represent GFP-labeled peroxisomes. (left) GFP-labeled peroxisomes are distributed in the cell body and along the length of processes. (middle) Plus end clusters are shown. DHC depletion allows kinesin-1 to transport GFP-labeled peroxisomes toward the tips of processes. (right) Minus end clusters are shown. KHC depletion allows dynein to transport GFP-labeled peroxisomes toward the cell center. Note that the altered morphology of KHC-depleted cells is discussed in Results. (B) Depletion of molecular motors does not alter peroxisome distribution along processes. Representative still images of S2 cells plated in the presence of CytoD on Con A–coated coverslips. (top) Differential interference contrast (DIC) images are shown. (bottom) Fluorescent images depicting GFP-labeled peroxisomes, corresponding to Videos 1–3. Arrowheads highlight the location of a single peroxisome within a process. Bar, 5 µm. (C) Bar graph representing the percentage of processes containing peroxisomes after RNAi-based depletion of motors (treatment with KHC, DHC, and Klp68D dsRNAs). Klp68D depletion served as a control. The total height of each column (blue and orange) represents the percentage of processes that contain peroxisomes anywhere along their length. The blue subcolumn represents the percentage of processes in which peroxisomes are limited to the shaft. The orange subcolumn represents the percentage of processes that contain at least one peroxisome at the tip. The schematic on the right identifies the parameters used to define shaft and tip (see Materials and methods). Data represent mean values ± SD from 120 cells per condition from three separate dsRNA treatments.

Kinesin-1 and cytoplasmic dynein function in an interdependent manner during peroxisome transport in S2 cells. (A) Potential outcomes of RNAi-based motor depletion if motors work independently of each other. Schematic depicts S2 cells plated in the presence of 1 µM CytoD to induce formation of microtubule-filled processes. Microtubule polarity is indicated with + and − signs. Green dots represent GFP-labeled peroxisomes. (left) GFP-labeled peroxisomes are distributed in the cell body and along the length of processes. (middle) Plus end clusters are shown. DHC depletion allows kinesin-1 to transport GFP-labeled peroxisomes toward the tips of processes. (right) Minus end clusters are shown. KHC depletion allows dynein to transport GFP-labeled peroxisomes toward the cell center. Note that the altered morphology of KHC-depleted cells is discussed in Results. (B) Depletion of molecular motors does not alter peroxisome distribution along processes. Representative still images of S2 cells plated in the presence of CytoD on Con A–coated coverslips. (top) Differential interference contrast (DIC) images are shown. (bottom) Fluorescent images depicting GFP-labeled peroxisomes, corresponding to Videos 1–3. Arrowheads highlight the location of a single peroxisome within a process. Bar, 5 µm. (C) Bar graph representing the percentage of processes containing peroxisomes after RNAi-based depletion of motors (treatment with KHC, DHC, and Klp68D dsRNAs). Klp68D depletion served as a control. The total height of each column (blue and orange) represents the percentage of processes that contain peroxisomes anywhere along their length. The blue subcolumn represents the percentage of processes in which peroxisomes are limited to the shaft. The orange subcolumn represents the percentage of processes that contain at least one peroxisome at the tip. The schematic on the right identifies the parameters used to define shaft and tip (see Materials and methods). Data represent mean values ± SD from 120 cells per condition from three separate dsRNA treatments.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal