Journal: Journal of Virology
Article Title: A recurrent adaptive mutation in the transmembrane 2B protein of an insect picorna-like virus in a nonnative host
doi: 10.1128/jvi.01239-25
Figure Lengend Snippet: Sting colocalizes with the CrPV 2B protein. ( A ) Confocal microscopy images of Drosophila S2 cells transfected with a combination of the indicated plasmids. Cells were fixed 48 h after transfection and stained with antibodies against the V5 tag and the cis Golgi protein GM130. ( B ) Violin plot of Pearson’s correlation coefficient (R) of Sting with eGFP ( n = 27), 2B-eGFP ( n = 22), or 2B(D29N)-eGFP ( n = 22) in individual cells, calculated in Fiji using the Costes method. The median is represented with a solid black line and the quartiles with dashed lines. Adjusted P -values from Tukey’s multiple comparison test are indicated with asterisks: ** P < 0.01, *** P < 0.001. ( C ) Co-immunoprecipitation of Sting with 2B and 2B(D29N). S2 cells were transfected with the indicated plasmids, and the cell lysates (input) and eGFP immunoprecipitates (eGFP ImmP) were analyzed by western blot using eGFP, V5, and GAPDH antibodies. The white arrow heads indicate Sting-V5 (expected size 43.1 kDa), black arrow heads indicate 2B-eGFP and 2B(D29N)-eGFP (43.3 kDa), and green arrow heads indicate eGFP. As previously reported for human STING, Drosophila Sting showed distinct migrating bands, which might be due to post-translational modifications . M, size marker.
Article Snippet: Membranes were stained with antibodies against GFP (rat, 1:1000, ChromoTek, 3h9-150), V5 tag (mouse, 1:2000, Invitrogen, 46-0705), or GAPDH (rabbit, 1:1000, Bioss, bs-8778R), followed by secondary antibodies IRDye 800CW goat anti-rat IgG (LICORBio, 926-32219), IRDye 800CW goat anti-mouse IgG (LICORBio, 926-32210), and IRDye 800CW goat anti-rabbit IgG (LICORBio, 926-32211), respectively.
Techniques: Confocal Microscopy, Transfection, Staining, Comparison, Immunoprecipitation, Western Blot, Marker