A.) Proposed mechanism for the removal of cell-surface CD54 mediated by TICTACs. B.) Depletion of cell-surface CD54 in M2-polarized RAW264.7 and BMA3.1A7 cells determined by live cell flow cytometry following 24 h of treatment with 25 nM unconjugated anti-CD54 antibody or conjugates. C.) Visualization of cell-surface CD54 by confocal microscopy after 25 nM TICTAC treatments for 24 h. D.) Dose-response curve for cell-surface CD54 removal in M2-polarized RAW264.7 cells following treatment with unconjugated antibody or 3b at 0.1 nM, 1 nM, 5 nM, 10 nM, and 100 nM for 24 h. E.) Time-course of CD54 downmodulation in M2-polarized RAW264.7 cells incubated with 25 nM unconjugated antibody or conjugates at 3 h, 8 h, 24 h, and 48 h. F.) Depletion of cell-surface CD54 in RAW264.7 and J774A.1 cells that have been polarized with IL-4 (25 ng/mL) or not following treatment with 25 nM unconjugated antibody or 3b.
For B and F, error bars represent the SD from 3 independent experiments. P values were determined by Welch’s two-tailed t-tests. Statistical significance was defined as P<0.05, and the asterisks * indicates P<0.1, ** indicates P<0.01, *** indicates P<0.001, and **** indicates P<0.0001.