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. 2012 Feb 3;7(2):e31004. doi: 10.1371/journal.pone.0031004

Figure 5. Chloroquine and hydroxhychloroquine inhibit CXCL12-mediated ERK phosphorylation.

Figure 5

(A) Immunostaining for phospho-ERK was performed for PANC-1, Hs-766T, AsPC-1, and MIAPaCa-2 cell lysates. Cells were pretreated with chloroquine or hydroxychloroquine (0.1 µM) for 30 minutes after which cells were exposed to CXCL12 (200 ng/ml) for 20 minutes. An antibody to total ERK1/2 was used as a loading control. The CXCL12-mediated increases in phospho-ERK were effectively abrogated with either chloroquine or hydroxychloroquine. (B) Pretreatment of PANC-1 and AsPC-1 cells with chloroquine and hydroxychloroquine (0.1–10 µM) for 5 minutes followed by exposure to CXCL12 (200 ng/ml) demonstrate dose-dependent effects of drug treatment on CXCL12-mediated ERK phosphorylation. (C) Western blots were scanned and quantified using the AlphaImager Tm3400 (Alpha Innotech). Fold changes for phospho-ERK compared to untreated controls were calculated as relative expression, which was normalized to protein band intensities of total ERK. The data shows the mean of triplicate experiments, with p-values<0.05 considered statistically significant.