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. 2019 Dec 4;5:148. doi: 10.1038/s41420-019-0229-8

Fig. 4. Induction of pro-inflammatory cytokine secretion in human RCC lines vs. normal HRPT cells is dependent on the mode of CD40 ligation.

Fig. 4

a–c RCC cell lines ACHN, 786-O and A-704, the positive control EJ and normal HRPT cells were treated with mCD40L. IL-8 (a), IL-6 (b) and GM-CSF (c) secretion in comparison to controls was detected at the indicated time-points post CD40 ligation (3, 6, 12 and 24 h) by ELISA assays (see Methods). Bars represent mean cytokine concentration (pg/ml) ± SEM for three technical replicates and are representative of two independent experiments. d, e RCC cell line 786-O (d) and normal HRPT cells (e) were treated with ‘mCD40L’ alongside negative controls (‘Control’), and CD40 was also activated with agonistic anti-CD40 mAb (‘G28-5′), and secretion of GM-CSF was measured by ELISA as above. Untreated RCC cells (‘786-O’ in d) and normal cells (‘HRPT’ in e), respectively, were also included as background controls. Bars show mean GM-CSF concentration (pg/ml) ± SEM for two technical replicates and are typical of two independent experiments.