Fig. 5

The combination of CBL0137 and rituximab exerted enhanced effects on B-NHL tumors in vitro and in vivo. A The synergy scores of CBL0137 and rituximab were calculated using the Bliss analysis. The concentration gradients of CBL0137 were 0.5, 1.0, 1.5, and 2.0 μM, and the concentration gradients of rituximab were 10, 20, 30, and 40 μg/mL. The combination responses of the two drugs were observed through the dose–response matrix. The 2D and 3D synergy maps above and below highlighted synergistic and antagonistic dose regions in red and green color respectively and showed the summary synergy scores of two drug combinations in three B-NHL cell lines. B LDH release assay of B-NHL cells treated with the indicated concentration of CBL0137 or rituximab and the combination of the two drugs for 24 h. C The Annexin V-FITC/PI staining results were shown on the left, which was that CBL0137 combined with rituximab has enhanced apoptotic effects on SU-DHL-4 and Raji cells. Statistical results of apoptosis assay were shown on the right, representing three independent experiments. (*P < 0.05, **P < 0.01, ***P < 0.001, Student’s t-test). D, E Representative western blotting results of B-NHL cells treated with the single or combination treatment for 24 h to determine changes in apoptosis-related proteins D and autophagy-related proteins E expression. F Experimental protocol of xenograft model of DLBCL. G SU-DHL-4 cells were subcutaneously implanted into male nude mice, and the mice were sacrificed after the treatment. The mice and their corresponding xenograft tumor specimens were photographed with a high-definition digital camera. H Tumor volume was measured every three days and tumor growth curves were plotted. ***P < 0.001, ****P < 0.0001. I Tumor weight in each group was measured and presented at the end of treatment. **P < 0.01, ***P < 0.001, ****P < 0.0001. J The body weight of mice in four groups (n = 8) was recorded weekly. K Histological examination of vital organs was performed by H&E staining. Scale bars = 50 μm. L Representative immunohistochemical images of Ki67, cleaved caspase-3, and LC3B were performed in the four groups of tumor samples. Scale bars = 50 μm. M, N Establishment of BL xenograft tumors. Raji cells were subcutaneously implanted into female nude mice, and the mice were sacrificed after the treatment. M Quantified results of tumor volume. N Quantified results of tumor weight