Panel A shows merged CD36WT, active beta 1 integrin, and fluorescence images with colocalization analysis and localization graph. The right panel is a barplot quantifying CD36WT colocalization with active beta-1-integrin, measured separately in cell center and periphery regions. The y-axis represents the colocalization extent, and the x-axis differentiates between center and periphery regions. Panel B shows a three-colored fixed cell image of CD36WT (magenta), beta-1-integrin (cyan), and actin (yellow) on the left, with CD36 particle detections overlaid on the actin channel in the middle. The right panel is a boxplot quantifying actin enrichment at CD36WT detections colocalized with or not colocalized with beta-1-integrin. The y-axis represents actin enrichment at CD36 detections, and the x-axis differentiates between colocalized and not colocalized detections. Panel C shows a three-colored fixed cell image of CD36WT (magenta), CD9 (cyan), and actin (yellow) on the left, with CD36 particle detections overlaid on the actin channel in the middle. The right panel is a boxplot quantifying actin enrichment at CD36WT detections colocalized with or not colocalized with CD9. The y-axis represents actin enrichment at CD36 detections, and the x-axis differentiates between colocalized and not colocalized detections.
CD36 colocalizes with activated β 1 -integrin; CD36 colocalized with β 1 -integrin or CD9 has higher actin enrichment around it than CD36 not colocalized with these interaction partners. (A) Representative two-color fixed cell image of CD36WT (magenta) and active β1-integrin (cyan; detected via the activation-specific antibody 12G10) in the left panel, and CD36 particle detections (shown as dots) overlaid on the CD36 channel in the middle panel. CD36WT colocalized with active β1-integrin is indicated by yellow/blue circles in the left/middle panel. Solid red lines show the segmented cell edge; dashed lines specify the boundary between center and periphery regions. Quantification of CD36WT colocalization with active β1-integrin, measured separately in cell center and periphery regions, is shown in the right panel. Right panel plot details are as in Fig. S2. Scale bar, 5 μm. N = 32 cells from four experimental repeats. (B and C) Left panels: Representative three-colored fixed cell images of CD36WT (magenta), β1-integrin (B) or CD9 (C) (both in cyan), and actin (yellow). Middle panels: CD36 particle detections (shown as dots) overlaid on the actin channel (same as that shown in left panels). Right panels: Quantification of actin enrichment measured at CD36WT detections colocalized with or not colocalized with β1-integrin (B) or CD9 (C). Solid red lines in image panels indicate ROI segmented based on actin intensity. In enrichment analysis plots (right panels), circles, red lines, shaded bars, and asterisks are as in Fig. 2. Scale bar, 5 μm. N = 32 cells from four experimental repeats.