Super‐resolution microscopy (SIM) imaging of Flag‐tagged proteins, cyclophilin A (CypA‐Flag), EB1‐Flag, or Flag‐CLIP170 bound to in vitro assembled CA‐NC complexes, stained with anti‐p24 AG3.0 (Red), anti‐Flag, or anti‐CLIP170 (Green). Scale bar, 2 μm. Representative images from different fields of view are shown.
WB analysis confirming protein expression in samples from (A) and (C).
Quantification (n = 3) of the association of each protein with 97 to 137 CA‐NC complexes, using untransfected lysates (Control) and EB1‐Flag as controls for non‐specific background staining.
Lysates from untransfected, CypA‐Flag-, or Flag‐CLIP170‐transfected cells were subjected to CA‐NC binding assays and WB analysis.
Levels of CypA or CLIP170 bound to WT or R18L CA‐NC assemblies in samples from (D) were quantified using Fiji (n = 4 experimental replicates; data are presented as a ratio of CypA or CLIP170 in bound versus input for each CA‐NC assembly, normalized to WT CA‐NC arbitrarily set to 1). Statistical significance was determined by t‐test (mean ± SEM). ***P < 0.001.
Data information: Molecular weight markers (in kDa) are shown to the right of WBs.
