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. 2021 Jan 1;11(7):3196–3212. doi: 10.7150/thno.51976

Figure 4.

Figure 4

Secreted activin A enhances PDAC stemness. (A-C) BxPC-3 cells were treated with indicated CM for 1 hour (A) or 48 hours (B-C) in the presence or absence of neutralizing antibody of activin A (4 μg/mL), and then Smad activation was measured by IB with the indicated antibodies (A), mRNA expressions of indicated CSC markers were analyzed by RT-qPCR (B), cancer cell survival in response to gemcitabine was determined by colony formation assay (C). (D) Luciferase activity assay in PDAC cells transfected with SBE4-Luc reporter vector for 24 hours, followed by treatment with or without activin A (10 ng/mL) for 16 hours. (E) IB analysis of Smad4 expression in 4 PDAC cell lines. (F) IB analysis of the indicated proteins in BxPC-3 or Mia-PaCa2 cells treated with activin A (10 ng/mL) for up to 8 hours. Data are presented as mean ± SD (n = 3) and are representative of at least three independent experiments. *P < 0.05, **P < 0.01 by two-tailed Student's t test. N.S., not significant.