Figure 8

TNFα induced transcriptional responses in AC16 cardiomyocytes result in corresponding changes in mRNA and protein levels with similar, but delayed, kinetics. A) Western blots of c-Fos, c-Jun, the NF-κB p50 subunit, and snRNP70 (a loading control) from whole cell extracts of control and TNFα-treated AC16 cells (25 ng/ml of TNFα for the indicated treatment times). The assays were repeated three times. B) Scatter plots showing the concentration of secreted cytokines determined using a Bio-rad Bio-Plex cytokine assay, including IL-6, IL-8, and MCP-1 as indicated in control and TNFα-treated AC16 cells (25 ng/ml of TNFα for the indicated treatment times). Each data point represents the mean ± SEM for three independent biological replicates. C) Scatter plots showing the level of transcription (by GRO-seq), mature mRNA (by RT-qPCR), and protein (by Western blotting or Bio-Plex cytokine assay) for FOS, JUN, NFKB1, IL6, IL8, and CCL2, as indicated, in control and TNFα-treated AC16 cells (25 ng/ml of TNFα for the indicated treatment times). Each data point represents the mean ± SEM for two (GRO-seq) or three (RT-qPCR, Western, Bio-Plex cytokine assay) independent biological replicates.