Fig. 5: Intracellular drug delivery using CLNCGTP/GTP*.
From: Reconstitution of microtubule into GTP-responsive nanocapsules
a Schematic illustration of the uptake of FITC-labeled CLNCGTP/GTP* into Hep3B cells. b Bright field (upper row) and CLSM images displaying FITC (middle row, green) in Hep3B cells and their merged images (lower row). The cells were incubated in EMEM containing CLNCGTP/GTP* (0.5 µg ml–1) for 2.5 h, rinsed with D-PBS, and further incubated in EMEM (10% FBS) for 1.5 h (i) and 21.5 h (ii). Scale bars, 20 µm. c Flow cytometry profiles showing FITC fluorescence of Hep3B cells (n > 660) incubated without (blue) and with FITC-labeled CLNCGTP/GTP* for 2.5 h, rinsed with D-PBS, and further incubated in EMEM (10% FBS) for 1.5 h (i, orange) and 21.5 h (ii, green). d Schematic illustration of the cellular uptake of CLNCGTP/GTP*⊃DOX. e Bright field (upper row) and CLSM images displaying DOX (middle row, red) in Hep3B cells and their merged images (lower row). The cells were incubated in EMEM containing CLNCGTP/GTP*⊃DOX ([CLNCGTP/GTP*] = 2.6 µg ml–1, [DOX] = 2 µM) for 2.5 h, rinsed with D-PBS, and further incubated in EMEM (10% FBS) for 1.5 h (iii) and 21.5 h (iv). Scale bars, 20 µm. f, g Flow cytometry profiles (f) showing DOX fluorescence of Hep3B cells (n > 390) and their normalized viabilities (g) determined using Cell Counting Kit-8 (n = 3). The cells were incubated without (blue) and with DOX (2 µM; orange), and CLNCGTP/GTP*⊃DOX ([CLNCGTP/GTP*] = 2.6 µg ml–1, [DOX] = 2 µM; red) for 2.5 h in EMEM, and then rinsed with D-PBS, followed by incubation in EMEM (10% FBS) for 21.5 h. Statistical significance was examined by two-sided Student’s t test (*p = 0.0094 < 0.01). Bars represent mean values ± SD from three different samples.
