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(A-B) <t>iSIM</t> images (see Methods) of PVD neurons (late L3 larvae) labeled with PVD::LifeAct::GFP (green) and PVD::mCherry::UNC-34 (magenta). Merge shown at top right. Insets depict punctate mCherry::UNC-34 in 1°, 2° and 3° PVD dendrites. (B) Note mCherry::UNC-34 puncta at tips (white arrowheads) of 3° dendrites. Scale bars are 5 μm. (C) Frequency (100%) at which tips of 3º dendrites show mCherry::UNC-34 puncta, ***p<0.001, Fischer’s exact test, N = 33 dendrite tips in 9 animals. Both iSIM and Airyscan images were used in this analysis. (D-E) Kymographs of PVD::mCherry::UNC-34 in PVD 3° dendrite captured in the iSIM <t>microscope.</t> Moving (green arrowheads) vs stationary (red arrowheads) mCherry::UNC-34 puncta are denoted. The distal end of the 3° dendrite is indicated by an asterisk and a red dashed arrow points toward the PVD cell soma. (F) Trafficking of mCherry::UNC-34 puncta in the PVD 3° dendrite (enclosed with rectangle of red dashed line) from 4E was analyzed using a spots tracking algorithm (Imaris) (see Methods). The net displacement (µm) of mobile mCh::UNC-34 puncta (N = 8) from position at t = 0 in a 3º dendrite plotted against time with a sampling interval of 2 s. Scale bar is 10 μm.
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(A-B) <t>iSIM</t> images (see Methods) of PVD neurons (late L3 larvae) labeled with PVD::LifeAct::GFP (green) and PVD::mCherry::UNC-34 (magenta). Merge shown at top right. Insets depict punctate mCherry::UNC-34 in 1°, 2° and 3° PVD dendrites. (B) Note mCherry::UNC-34 puncta at tips (white arrowheads) of 3° dendrites. Scale bars are 5 μm. (C) Frequency (100%) at which tips of 3º dendrites show mCherry::UNC-34 puncta, ***p<0.001, Fischer’s exact test, N = 33 dendrite tips in 9 animals. Both iSIM and Airyscan images were used in this analysis. (D-E) Kymographs of PVD::mCherry::UNC-34 in PVD 3° dendrite captured in the iSIM <t>microscope.</t> Moving (green arrowheads) vs stationary (red arrowheads) mCherry::UNC-34 puncta are denoted. The distal end of the 3° dendrite is indicated by an asterisk and a red dashed arrow points toward the PVD cell soma. (F) Trafficking of mCherry::UNC-34 puncta in the PVD 3° dendrite (enclosed with rectangle of red dashed line) from 4E was analyzed using a spots tracking algorithm (Imaris) (see Methods). The net displacement (µm) of mobile mCh::UNC-34 puncta (N = 8) from position at t = 0 in a 3º dendrite plotted against time with a sampling interval of 2 s. Scale bar is 10 μm.
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(A-B) <t>iSIM</t> images (see Methods) of PVD neurons (late L3 larvae) labeled with PVD::LifeAct::GFP (green) and PVD::mCherry::UNC-34 (magenta). Merge shown at top right. Insets depict punctate mCherry::UNC-34 in 1°, 2° and 3° PVD dendrites. (B) Note mCherry::UNC-34 puncta at tips (white arrowheads) of 3° dendrites. Scale bars are 5 μm. (C) Frequency (100%) at which tips of 3º dendrites show mCherry::UNC-34 puncta, ***p<0.001, Fischer’s exact test, N = 33 dendrite tips in 9 animals. Both iSIM and Airyscan images were used in this analysis. (D-E) Kymographs of PVD::mCherry::UNC-34 in PVD 3° dendrite captured in the iSIM <t>microscope.</t> Moving (green arrowheads) vs stationary (red arrowheads) mCherry::UNC-34 puncta are denoted. The distal end of the 3° dendrite is indicated by an asterisk and a red dashed arrow points toward the PVD cell soma. (F) Trafficking of mCherry::UNC-34 puncta in the PVD 3° dendrite (enclosed with rectangle of red dashed line) from 4E was analyzed using a spots tracking algorithm (Imaris) (see Methods). The net displacement (µm) of mobile mCh::UNC-34 puncta (N = 8) from position at t = 0 in a 3º dendrite plotted against time with a sampling interval of 2 s. Scale bar is 10 μm.
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(A-B) <t>iSIM</t> images (see Methods) of PVD neurons (late L3 larvae) labeled with PVD::LifeAct::GFP (green) and PVD::mCherry::UNC-34 (magenta). Merge shown at top right. Insets depict punctate mCherry::UNC-34 in 1°, 2° and 3° PVD dendrites. (B) Note mCherry::UNC-34 puncta at tips (white arrowheads) of 3° dendrites. Scale bars are 5 μm. (C) Frequency (100%) at which tips of 3º dendrites show mCherry::UNC-34 puncta, ***p<0.001, Fischer’s exact test, N = 33 dendrite tips in 9 animals. Both iSIM and Airyscan images were used in this analysis. (D-E) Kymographs of PVD::mCherry::UNC-34 in PVD 3° dendrite captured in the iSIM <t>microscope.</t> Moving (green arrowheads) vs stationary (red arrowheads) mCherry::UNC-34 puncta are denoted. The distal end of the 3° dendrite is indicated by an asterisk and a red dashed arrow points toward the PVD cell soma. (F) Trafficking of mCherry::UNC-34 puncta in the PVD 3° dendrite (enclosed with rectangle of red dashed line) from 4E was analyzed using a spots tracking algorithm (Imaris) (see Methods). The net displacement (µm) of mobile mCh::UNC-34 puncta (N = 8) from position at t = 0 in a 3º dendrite plotted against time with a sampling interval of 2 s. Scale bar is 10 μm.
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(A-B) <t>iSIM</t> images (see Methods) of PVD neurons (late L3 larvae) labeled with PVD::LifeAct::GFP (green) and PVD::mCherry::UNC-34 (magenta). Merge shown at top right. Insets depict punctate mCherry::UNC-34 in 1°, 2° and 3° PVD dendrites. (B) Note mCherry::UNC-34 puncta at tips (white arrowheads) of 3° dendrites. Scale bars are 5 μm. (C) Frequency (100%) at which tips of 3º dendrites show mCherry::UNC-34 puncta, ***p<0.001, Fischer’s exact test, N = 33 dendrite tips in 9 animals. Both iSIM and Airyscan images were used in this analysis. (D-E) Kymographs of PVD::mCherry::UNC-34 in PVD 3° dendrite captured in the iSIM <t>microscope.</t> Moving (green arrowheads) vs stationary (red arrowheads) mCherry::UNC-34 puncta are denoted. The distal end of the 3° dendrite is indicated by an asterisk and a red dashed arrow points toward the PVD cell soma. (F) Trafficking of mCherry::UNC-34 puncta in the PVD 3° dendrite (enclosed with rectangle of red dashed line) from 4E was analyzed using a spots tracking algorithm (Imaris) (see Methods). The net displacement (µm) of mobile mCh::UNC-34 puncta (N = 8) from position at t = 0 in a 3º dendrite plotted against time with a sampling interval of 2 s. Scale bar is 10 μm.
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Image Search Results


(A-B) iSIM images (see Methods) of PVD neurons (late L3 larvae) labeled with PVD::LifeAct::GFP (green) and PVD::mCherry::UNC-34 (magenta). Merge shown at top right. Insets depict punctate mCherry::UNC-34 in 1°, 2° and 3° PVD dendrites. (B) Note mCherry::UNC-34 puncta at tips (white arrowheads) of 3° dendrites. Scale bars are 5 μm. (C) Frequency (100%) at which tips of 3º dendrites show mCherry::UNC-34 puncta, ***p<0.001, Fischer’s exact test, N = 33 dendrite tips in 9 animals. Both iSIM and Airyscan images were used in this analysis. (D-E) Kymographs of PVD::mCherry::UNC-34 in PVD 3° dendrite captured in the iSIM microscope. Moving (green arrowheads) vs stationary (red arrowheads) mCherry::UNC-34 puncta are denoted. The distal end of the 3° dendrite is indicated by an asterisk and a red dashed arrow points toward the PVD cell soma. (F) Trafficking of mCherry::UNC-34 puncta in the PVD 3° dendrite (enclosed with rectangle of red dashed line) from 4E was analyzed using a spots tracking algorithm (Imaris) (see Methods). The net displacement (µm) of mobile mCh::UNC-34 puncta (N = 8) from position at t = 0 in a 3º dendrite plotted against time with a sampling interval of 2 s. Scale bar is 10 μm.

Journal: bioRxiv

Article Title: Actin assembly and non-muscle myosin activity drive dendrite retraction in an UNC-6/Netrin dependent self-avoidance response

doi: 10.1101/492769

Figure Lengend Snippet: (A-B) iSIM images (see Methods) of PVD neurons (late L3 larvae) labeled with PVD::LifeAct::GFP (green) and PVD::mCherry::UNC-34 (magenta). Merge shown at top right. Insets depict punctate mCherry::UNC-34 in 1°, 2° and 3° PVD dendrites. (B) Note mCherry::UNC-34 puncta at tips (white arrowheads) of 3° dendrites. Scale bars are 5 μm. (C) Frequency (100%) at which tips of 3º dendrites show mCherry::UNC-34 puncta, ***p<0.001, Fischer’s exact test, N = 33 dendrite tips in 9 animals. Both iSIM and Airyscan images were used in this analysis. (D-E) Kymographs of PVD::mCherry::UNC-34 in PVD 3° dendrite captured in the iSIM microscope. Moving (green arrowheads) vs stationary (red arrowheads) mCherry::UNC-34 puncta are denoted. The distal end of the 3° dendrite is indicated by an asterisk and a red dashed arrow points toward the PVD cell soma. (F) Trafficking of mCherry::UNC-34 puncta in the PVD 3° dendrite (enclosed with rectangle of red dashed line) from 4E was analyzed using a spots tracking algorithm (Imaris) (see Methods). The net displacement (µm) of mobile mCh::UNC-34 puncta (N = 8) from position at t = 0 in a 3º dendrite plotted against time with a sampling interval of 2 s. Scale bar is 10 μm.

Article Snippet: Time-lapse movies from the iSIM microscope were deconvolved with metamorph.

Techniques: Labeling, Microscopy, Sampling