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. Author manuscript; available in PMC: 2020 Aug 4.
Published in final edited form as: Cancer Discov. 2019 Aug 21;9(11):1574–1589. doi: 10.1158/2159-8290.CD-18-1308

Figure 2: ADAMDEC1 is associated with GBM stemness and secreted by GSCs.

Figure 2:

(A) ADAMDEC1 protein is expressed in GSC, but not in NSTC culture paradigms. Likewise, GSCs secrete ADAMDEC1 into the medium. Depicted are Western blots from cell culture conditioned medium, with 10 µg protein lysate loaded per lane. (B) Knockdown of ADAMDEC1 using shRNA. Compared to non-targeting (NT) constructs, ADAMDEC1 knockdown results in decreased SOX2 and increased GFAP expression. (C) Sphere-forming frequency is reduced after ADAMDEC1 knockdown (data from two independent experiments, one-way ANOVA). (D) ADAMDEC1 knockdown results in decreased cell proliferation in GSC cultures (n=6, non-linear regression). (E) Orthotopic implantation of ADAMDEC1 knockdown cells significantly increases survival of tumor-bearing animals compared to control cells (median survival NT=43 d, #4 and #5=100d; n=10 mice/group; log rank test). (F) Treatment of GSCs with recombinant ADAMDEC1 results in increased levels of FGF2, but not GRO alpha, in the culture supernatant in a concentration-dependent manner (n=3, two-way ANOVA with Dunnet post-test). (G) ELISA shows increased levels of FGF2 in ADAMDEC1-treated GSC, but not in NSTC cultures (data from two independent experiments). (H) Western blot depicting FGFR phosphorylation after knockdown of ADAMDEC1, or after treatment with rADAMDEC1.