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. 2019 Oct 24;20(21):5287. doi: 10.3390/ijms20215287

Figure 4.

Figure 4

Signaling molecules involved in PDT-triggered monocyte migration. (A) Transwell migration assay of monocytes in response to the indicated treatments. Monocytes pretreated for 1 h at 37 °C with the inhibitors LY294002 (25 μM), wortmannin (100 nM), Akt inhibitor VIII (1 μM), AG490 (100 μM), PD98059 (10 μM) and staurosporine (1 nM) were stimulated for 90 min at 37 °C with PBS (NT) or PDT (0.5 µg/mL). Bars represent the means ± SD of 3 independent experiments performed in triplicate. Statistical analysis was performed by one-way ANOVA and the Bonferroni’s post-test was used to compare data, *** p < 0.001. (B) Monocytes were stimulated with 5 µg/mL of PDT at 37 °C for the indicated times. Not treated cells (NT) were used as control (lane 1). Western blot analysis of monocyte lysates shows that PDT activates Akt, as shown by the respective phosphorylation state, verified by densitometric analysis and plotting of the phospho-Akt/total Akt (pAkt/tAkt). In the left panel blots from one representative experiment of three with similar results are shown. In the right panel, values reported for Akt phosphorylation are the mean ± SD of three independent experiments. Statistical analysis was performed by one-way ANOVA and the Bonferroni’s post-test was used to compare data, * p < 0.05.