Figure 4

SLB mediated D6F10 delivery in HeLa cells.

Confocal images of HeLa cells treated with (A) D6F10 633, (B) SLB 488-D6F10 633 and (C) CP 488-D6F10 633 for 4 hours. Confocal images of HeLa cells treated with SLB 488-D6F10 633 in the absence (D) and in the presence (E) of sheep serum (40 ng/μl). Green = SLB 488/CP 488, red = D6F10 633, blue = DAPI stained nuclei and Merge = all three fluorophores. (F) Bar diagram representing translation assay of HeLa cells treated with CP (1.5 nM), SLB (1.5 nM), abrin (0.166 nM), abrin-D6F10 (1:25, 1:50), SLB-D6F10 (10:25, 10:50) and CP. For the latter three samples, after pre-treatment of cells with SLB-D6F10 and CP-D6F10 for 2 hours, cells were treated with abrin (0.166 nM) for 7 hours. Total counts per minute (cpm) of tritium is represented on the Y axis. (G) Percentage of dead population obtained by cell cycle progression analysis of HeLa cells treated with CP (10 nM), SLB (10 nM), abrin (0.1 nM), abrin-D6F10 (1:50) for 36 hours and SLB-D6F10 (1:50) for 4 hours followed by abrin for 36 hours. The propidium iodide stained cells were analysed by BD FACS. ANOVA followed by Tukey’s multiple comparison tests was performed. Ns = not significant, ***p < 0.05.