Differential expression of TNFα and hsa-miR-34c in human donor PBMCs following exposure to wild-type (wt) HCT116 or HMGB1 stable knock-down (kd) lysates.

<p><b>A</b>: HCT116 cells were stably transfected with a shHMGB1 vector in the presence of Puromycin (100 ug/ml). The clone with complete knockdown of HMGB1 (indicated by the arrow) was chosen to make necrotic lysates. <b>B</b>: TNFα ELISA showing differential release of TNFα in human donor PBMC cell cultures following exposure to HCT116 lysates (wild-type, wt or HMGB1 knockdown, kd) for 24 hrs. The amount of HCT116 lysates used was 104 cells/ml of human PBMC culture. Data shown are the average±s.d. of three independent experiments, each from a different donor, measured in triplicates, where **indicates p<0.01, by Student’s t test. <b>C</b>: Fold changes in expression (as log-2-transformed RQ values) of hsa-miR-34c and hsa-miR-214 in donor PBMCs exposed to the indicated HCT116 necrotic lysates for 48 hrs. Values were measured using TaqMan real-time RT-PCR. Data shown are the average±s.d. of three independent experiments, each from an individual donor, measured in triplicates, where **indicates p<0.01, and where ***indicates p<0.001, by paired Student’s t test.