Fig. 5

Ixa + Dina combination demonstrates increased in vitro efficacy against HCC PDXOs. A Immunoblotting of ER stress, NFκB and cell cycle proteins in representative PDXO lines, PDXO1 and 11. B Brightfield images of HCC PDXOs treated with vehicle (DMSO), ixazomib, dinaciclib or Ixa + Dina. Scale bar = 100 μm. C Average viability of PDXOs when treated with ixazomib (0.1 μM) and dinaciclib (0.01 μM) as monotherapy and in combination. Data presented as mean ± SD, n = 5. *, P < 0.05; **, P < 0.01; ***, P < 0.001. D Histograms of cell cycle analysis and the corresponding quantification of the proportion of cells in A (apoptotic), G0/G1, S, and G2/M phases of the cell cycle after 24 h drug treatment in PDXO11. Data shown as mean ± SD, n = 3. *, P < 0.05; **, P < 0.01; ***, P < 0.001, compared to Ixa + Dina. Black and grey asterisks indicate comparison for G2/M and apoptotic phases, respectively. E Quantification of annexin V/PI-stained PDXO1 and PDXO11 after Ixa + Dina treatment for 24 hours. Data presented as mean ± SD, n = 3. **, P < 0.01; ***, P < 0.001, compared to Ixa + Dina. Black and grey asterisks indicate comparison for early and late apoptosis groups, respectively. F Immunoblotting of apoptotic markers after treatment with Ixa + Dina in HCC PDXOs. All statistical analyses were performed using two-tailed Student’s t test