Figure 1
Figure 1. Localization and dynamics of MT1-MMP-mCherry in macrophages. (A-B) Still images of confocal time-lapse videos (supplemental Videos 1-2) of a macrophage expressing MT1-MMP–mCherry. Note the central accumulation and also vesicles throughout the cytoplasm of a quiescent (A), or at the leading edge of a randomly migrating cell (B). (C-F) Micrographs of a primary macrophage (C) expressing MT1-MMP–mCherry, (D) stained for trans-Golgi marker TGN46 using specific primary and Alexa 488–labeled secondary antibody, and (E) stained for cis-Golgi marker GM130 using specific primary and Cy5-labeled secondary antibody. (F) Merge of images in panels C, D, and E. White bar indicates 10 μm, white line in panel C indicates axis used for the acquisition of respective xz side views shown in panels Ci, Di, Ei, and Fi. (G-H) MT1-MMP–mCherry vesicles move bidirectionally along microtubules. (G) Still image from confocal time-lapse video (supplemental Videos 3-4) of a macrophage expressing MT1-MMP–mCherry and α-tubulin–GFP. White scale bar indicates 10 μm; white box indicates detail area shown in panel H. Detail images (supplemental Video 4) showing movement of MT1-MMP–mCherry vesicles along microtubules. (H left panel) Merged images; (middle panel) MT1-MMP–mCherry signal; (right panel) α-tubulin–GFP signal. Block of images on left shows anterograde movement of a vesicle toward the cell periphery; block of images on right shows retrograde movement of the same vesicle toward the cell center. Time in seconds since the start of the experiment is indicated on the right. (I-J) Movement of MT1-MMP–mCherry vesicles along actin cables in the cell periphery. Confocal micrograph of macrophage expressing MT1-MMP–mCherry and Lifeact-GFP, labeling F-actin. White scale bar indicates 10 μm. White box indicates area shown in detail images in panel J. Detail images from time-lapse video (supplemental Video 5) showing bidirectional movement of MT1-MMP–mCherry along actin cables. Elapsed time between frames is indicated below. White arrows indicate direction of MT1-MMP–mCherry movement. White bar indicates 5 μm. (K) Velocities of MT1-MMP–mCherry vesicles. Box plot diagram shows velocities in micrometers per second for vesicles moving through the central parts of the cell (nonperipheral; n = 20) and for vesicles moving within the cell periphery (within 5 μm of the cell border; peripheral; n = 20). Box plots show mean values with a statistical cut off of ± 25%, with respective SD.

Localization and dynamics of MT1-MMP-mCherry in macrophages. (A-B) Still images of confocal time-lapse videos (supplemental Videos 1-2) of a macrophage expressing MT1-MMP–mCherry. Note the central accumulation and also vesicles throughout the cytoplasm of a quiescent (A), or at the leading edge of a randomly migrating cell (B). (C-F) Micrographs of a primary macrophage (C) expressing MT1-MMP–mCherry, (D) stained for trans-Golgi marker TGN46 using specific primary and Alexa 488–labeled secondary antibody, and (E) stained for cis-Golgi marker GM130 using specific primary and Cy5-labeled secondary antibody. (F) Merge of images in panels C, D, and E. White bar indicates 10 μm, white line in panel C indicates axis used for the acquisition of respective xz side views shown in panels Ci, Di, Ei, and Fi. (G-H) MT1-MMP–mCherry vesicles move bidirectionally along microtubules. (G) Still image from confocal time-lapse video (supplemental Videos 3-4) of a macrophage expressing MT1-MMP–mCherry and α-tubulin–GFP. White scale bar indicates 10 μm; white box indicates detail area shown in panel H. Detail images (supplemental Video 4) showing movement of MT1-MMP–mCherry vesicles along microtubules. (H left panel) Merged images; (middle panel) MT1-MMP–mCherry signal; (right panel) α-tubulin–GFP signal. Block of images on left shows anterograde movement of a vesicle toward the cell periphery; block of images on right shows retrograde movement of the same vesicle toward the cell center. Time in seconds since the start of the experiment is indicated on the right. (I-J) Movement of MT1-MMP–mCherry vesicles along actin cables in the cell periphery. Confocal micrograph of macrophage expressing MT1-MMP–mCherry and Lifeact-GFP, labeling F-actin. White scale bar indicates 10 μm. White box indicates area shown in detail images in panel J. Detail images from time-lapse video (supplemental Video 5) showing bidirectional movement of MT1-MMP–mCherry along actin cables. Elapsed time between frames is indicated below. White arrows indicate direction of MT1-MMP–mCherry movement. White bar indicates 5 μm. (K) Velocities of MT1-MMP–mCherry vesicles. Box plot diagram shows velocities in micrometers per second for vesicles moving through the central parts of the cell (nonperipheral; n = 20) and for vesicles moving within the cell periphery (within 5 μm of the cell border; peripheral; n = 20). Box plots show mean values with a statistical cut off of ± 25%, with respective SD.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal