Extended Data Fig. 2: Subcellularly targeted HaloAKAR biosensors.
a,b, Plasma membrane targeting via fusion to the CAAX sequence of KRAS (a) or Lyn11 (b). c, Outer mitochondrial membrane targeting by fusing to Tomm20. d, Actin targeting via fusion with β-actin. e, Microtubule targeting via microtubule binding protein CEP41. f, Clathrin targeting via fusion with clathrin light chain. Average time courses of HeLa cells expressing targeted HaloAKAR2.0, HaloAKAR2.1, or HaloAKAR2.2 labeled with JF635-CA and stimulated with 50 μM Fsk and 100 μM IBMX (Fsk/IBMX) are shown along with representative images after stimulation. Time courses and images are representative of three replicates. Thick and thin solid lines indicate mean and single-cell responses, respectively; shaded areas correspond to standard deviation (a: n = 8, 8, 7 cells; b: n = 7, 6, 4 cells; c: n = 7, 7, 5 cells; d: n = 5, 7, 4 cells; e: n = 6, 4, 5 cells; f: n = 5, 8, 6 cells). Scale bars, 20 µm.
