Formation of ER–FA contacts is dependent on the binding of Rab18 and KIF5B to KNT1. (a) U2OS cells silenced with siRNA control or siRNA targeting Rab18 were cotransfected with ER-GFP and RFP-vinculin. The images represent maximum-intensity projections from Z-stacks. The insets show magnifications of the boxed regions. Scale bar: 10 µm; insets: 2 µm. (b) Quantification of the percentage of FAs in contact with the ER. The graph represents the mean ± SEM from three independent experiments (n > 30 cells). ***, P < 0.001. (c) U2OS cells were treated with either control siRNA or siRNA directed against KNT1 and transfected with either dsRed-ER alone or together with either GFP KNT1 FL, TM, or Δ201–445. Cells were fixed and stained with anti-vinculin antibody. The images represent maximum-intensity projections from Z-stacks. The insets show magnification of boxed regions to illustrate contact points between ER and FAs. Scale bars: 10 µm; insets: 2 µm. (d) Quantification of the percentage of FAs in contact with the ER. The graph represents the mean ± SEM from three independent experiments (n > 30 cells). *, P < 0.05; ***, P ≤ 0.001; n.s., not significant.