Fig. 4
MSC-CMSTIM induced ERK1/2 phosphorylation is mediated via (trans)activation of EGFR. a: 1 h pre-incubation of NCI-H292 with 0.05 μM PF04217903 and/or 10 μM AG1478, followed by 15 min stimulation with MSC-CMSTIM or DMEMSTIM, or DMEMCTRL showed that inhibition of EGFR decreased ERK1/2 phosphorylation as determined using Western blot. b: Densitometry for Fig. 4a. Error bars represent SEM, n = 5; (*) p < 0.05. c: 1 h pre-incubation with 2 μg/ml neutralizing anti-EGFR antibodies and 10 μM TAPI-1 followed by incubation with MSC-CMSTIM during 15 min showed that these agents decreased ERK1/2 phosphorylation. d: Densitometry for Fig. 4c. Error bars represent SEM, n = 3; (*) p < 0.05. e: NCI-H292 cells were incubated with MSC-CMSTIM or DMEMSTIM and harvested after 9 h for mRNA analysis. mRNA expression of the EGFR ligands AREG, HBEGF and TGFA was determined by qPCR. MSC-CMSTIM significantly increased the expression of AREG and HBEGF. Expression of TGFA increased but this was not significant. f: mRNA expression of the proliferation marker CCDN1 was significantly increased. Values were normalized against RPL13A and ATP5B reference genes. Box and whiskers represent median, interquartile range and minimum and maximum. n = 4; (*) p < 0.05