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. 2019 May 29;11(6):745. doi: 10.3390/cancers11060745

Figure 8.

Figure 8

HMF-mediated proliferation boost of PancTu-I cells is VEGF dependent. (A) RNA was isolated from HSC or HMF after 6 days of coculture with PDAC cells and mRNA levels of murine VEGF-A were analyzed by qRT-PCR. VEGF-A gene expression was normalized to GAPDH expression and normalized expression of HSC cocultured with PancTu-I shCtrl cells was set as 1. For blocking experiments, PancTu-I shCtrl or PancTu-I shTR2 cells were indirectly cocultured in the presence of HMF and treated with 10 µg/mL of either Rituximab as control or Aflibercept. After 6 days of coculture, (B) vital cell count and (C) the proportion of Ki67+ cells were determined. The percentage of proliferating cells was determined by immunofluorescence Ki67-Alexa-488 staining. Data are normalized to PancTu-I shCtrl cells + HMF treated with Rituximab. (D) Representative images of immunofluorescent Ki67 staining are shown. Scale Bars: 100 µm. Data represent the mean ± SEM of 4–5 independent experiments; * = p < 0.05.