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Fig. 6 | Journal of Experimental & Clinical Cancer Research

Fig. 6

From: Combination of Enzastaurin and Ibrutinib synergistically induces anti-tumor effects in diffuse large B cell lymphoma

Fig. 6

Whole-transcriptome changes in DLBCL occur in response to the combination of enzastaurin and ibrutinib. HBL-1 and TMD8 cells were exposed for 24 h with enzastaurin (HBL-1 2 μM, TMD8 4 μM) and/or ibrutinib (HBL-1 0.02 μM, TMD8 0.003 μM). RNA was collected for RNA sequencing. a Venn diagram illustrating the number of overlapping downregulated gene between different groups. b Significantly down-regulated genes from top ranked pathways (by KEGG) are represented in the heatmap. Colors scale bar represents from higher (red) to lower (blue) expression. Gene expression levels are expressed in FPKM values, differences shown in color scale after Z-score transformation. Down-regulated genes were determined by log2foldchange<0. FPKM, fragments per kilo base of exon per million fragments mapped. c The expressions of NOTCH1 in DLBCL cell lines were detected using qRT-PCR and western blot. d NOTCH1 knockdown by shRNA was validated by western blot in HBL-1, TMD8, OCI-LY7, SU-DHL-6 cells. β-actin is shown as a loading control. e The expressions of NOTCH1 gene were further confirmed in DLBCL cells after pre-treated with enzastaurin and/or ibrutinib. f The DLBCL cells were transfected with shRNA targeting NOTCH1, or treated with enzastaurin and ibrutinib for 48 h, 72 h. The cell viability of tumor cells was determined using the Cell Titer-Glo luminescent cell viability assay. Results are expressed as mean ± SD, data are representative of three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001 compared with control group; # p < 0.05, ## p < 0.01 compared with enzastaurin group

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