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. 2017 Jun 13;7:3420. doi: 10.1038/s41598-017-03744-3

Figure 3.

Figure 3

hrSerpinB3-dependent induction of oriented migration relies on intracellular generation of ROS and specific signaling pathways. (a,b) Confluent and 24 hr starved LX2 cells were left untreated or exposed to either hrSerpinB3 (100 ng/ml) or PDGF-BB (10 ng/ml, positive control) and then evaluated for early (i.e., 15 minutes) intracellular generation of ROS as shown by DCFH-DA fluorescence (a) or flow cytometry analysis (b). (c,d) Analysis of oriented migration in the modified Boyden’s chamber assay (c) or Western blot analysis of activation of JNK1/2 and Akt signaling pathways (d) of control LX2 cells or LX2 cells exposed to hrSerpinB3 (100 ng/ml) in the presence of pharmacological inhibitors of JNK1/2 (SP, SP600125,) and Akt (LY, LY294002) or in the presence of apocynin (APO) or diphenylen-iodonium (DPI), all added 30 min before addition of hrSerpinB3. Data in bar graphs are expressed as means ± SEM of three independent experiments (*p < 0.05 or **p < 0.01 vs control values, #p < 0.05 vs related stimulus). Images from Western blot analysis or from evaluation of ROS generation are representative from at least three experiments performed. The cropped gels shows in this Fig. have been run under the same experimental conditions.