Visualizing AP2M1 and Influenza A NP Cotrafficking

Live imaging was used to visualize the cotrafficking of AP2M1 and influenza A NP according to a previous report with some modifications (37 (link)). Time-lapse images were taken using the Nikon Ti2-E Widefield Microscope with a 100× 1.46 oil objective in a heated (37°C) chamber. GFP-labeled NP protein and mCherry-fused AP2M1 protein were tracked by sequential imaging every 2 s with 50- and 200-ms exposures for each channel, respectively. Individual colocalized puncta run lengths and transport velocities were calculated using the Track Points for MetaMorph analysis software, measuring the distance traveled (in any direction) between frames for each respective puncta. The movies were made using the Stack function for MetaMorph analysis software via accumulating the relative frames in order.

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Yuan S., Chu H., Huang J., Zhao X., Ye Z.W., Lai P.M., Wen L., Cai J.P., Mo Y., Cao J., Liang R., Poon V.K., Sze K.H., Zhou J., To K.K., Chen Z., Chen H., Jin D.Y., Chan J.F, & Yuen K.Y. (2020). Viruses harness YxxØ motif to interact with host AP2M1 for replication: A vulnerable broad-spectrum antiviral target. Science Advances, 6(35), eaba7910.

Publication 2020

Ti2-E Widefield Microscope

Frames Influenza Microscope Protein

Corresponding Organization : Hainan Medical University

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Variable analysis

independent variables

Live imaging

dependent variables

Cotrafficking of AP2M1 and influenza A NP
Colocalized puncta run lengths
Transport velocities

control variables

Heated chamber (37°C)
Sequential imaging every 2 s with 50- and 200-ms exposures for each channel, respectively
Nikon Ti2-E Widefield Microscope with a 100× 1.46 oil objective

controls

Previous report with some modifications (37 (link))

Annotations

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