PLA indicates that CD36 forms proximity associations with β ₁ -integrin that are reduced by the G12V mutation. (A) Quantification of PLA signal from spinning disk confocal imaging, representing the extent of interaction between the antibody pairs against the proteins indicated below the x-axis throughout the whole cell. The boxplot for each pair displays the percentage of the cell area occupied by the PLA puncta for that pair. Nonparametric ANOVA Kruskal–Wallis test followed by a two-sided Wilcoxon rank-sum test were employed to compare the different conditions, with ** and *** indicating P ≤ 0.01 and 0.001, respectively. N = 233 (CD36^WT vs. K20), 120 (CD36^G12V vs. K20), 12 (CD36^WT vs. 12G10), 71 (CD36^G12V vs. 12G10), 12 (CD36^WT vs. mitofilin), and 43 (CD36^G12V vs. mitofilin) from two to three experimental repeats per pair. (B) Quantification of PLA signal from TIRFM, representing the extent of interaction between the antibody pairs against the proteins indicated below the x-axis on the cell surface. The extent of interaction is expressed as the ratio of PLA signal detection density in CD36-expressing cells to PLA signal detection density in cells not expressing CD36. Circles, red lines, shaded bars, and * are as in Fig. 2. N = 24–25 cells for each pair, combined from 3 experimental repeats. (C–H) Representative fixed cell images showing PLA signals acquired via TIRFM (corresponding to the quantifications shown in B) for the indicated protein pairs in transfected cells expressing CD36 (C, E, and G) or not expressing CD36 (D, F, and H). Left panels show PLA signal alone, while right panels show merged images of CD36 (magenta), actin (phalloidin, cyan), and PLA puncta (yellow). Solid red line indicates segmented cell edge. Scale bar (in all images), 5 μm.