Skip to main content
. 2020 Feb 5;9(2):11. doi: 10.1038/s41389-020-0199-y

Fig. 5. Dasatinib (Src inhibitor) inhibits spreading of EGFRvIII/EGFRwt multicellular cultures.

Fig. 5

a We hypothesize that pSrc inhibition, using, for example, dasatinib at non-toxic concentrations, may reduce the spreading of EGFRvIII/EGFRwt CC. b Western blot analysis shows that Src activity was inhibited using non-toxic concentration (100–200 nM) of dasatinib in CC after 24 h of treatment. WB images are representative of at least three independent experiments. c Examination of CC survival, treated with a range of dasatinib concentrations, for 72 h. Bars represent % of viable cells as assessed by Methylene blue assay (error bars represent SE). d CC was treated for 24 h with 100 nM of dasatinib or 100 nM erlotinib. Hoechst-labeled cell nuclei were imaged using live-cell imaging chambers (Eclipse Ti-E, Nikon inverted microscope Scale bars, 100 μm, ×10 lens). e Probability of finding a pair of the U87EGFRwt cells at r (0–200 μm) was calculated at 0 and 72 h as described in Methods. Plots represent the distribution of cell–cell separation distances in CC treated with dasatinib, with erlotinib or in the untreated CC. *P ≤ 0.03 was found for CC treated with dasatinib vs. untreated and for CC treated with dasatinib vs erlotinib. f, g CC (Control) and CC treated with dasatinib for 18 h were imaged using ×10 lens and live-cell imaging chambers. Cell velocity, V (f) and the line length of cell migration (g) were calculated using the NIS-Elements (Nikon); *P < 0.001. Data were collected from eight fields (~200 cells/field). See also video 2(A and B). h, i mCherry-labeled U87EGFRwt cells were transfected with 0.5 nM Src siRNA or negative control (NC) siRNA. Non-transfected (indicated as control) and siRNA transfected U87EGFRwt cells were co-cultured with U87EGFRvIII cells (CC), in ratio of 9:1. 72 h after seeding, cells were imaged, and the probability of finding a pair of the U87EGFRwt cells at r (0–200 μm) was calculated as described in Methods. Plots represent the distribution of cell–cell separation distances in untreated CC or CC transfected with 0.5 nM Src/NC siRNA. *P < 0.001, **P = 0.01 (for CC treated with Src siRNA vs untreated). Data were collected from eight fields (~200 cells/field).