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. Author manuscript; available in PMC: 2021 Jun 7.
Published in final edited form as: Oncogene. 2020 Dec 7;40(4):848–862. doi: 10.1038/s41388-020-01564-w

Fig.6. Afatinib decreased the CSC population via EGFR/ERK/FOXA2/SOX9 signaling in PCSCs.

Fig.6.

a PCR array for CSC and self-renewal markers in SP/CSC cells (SP-SW1990) after 4 days of treatment with afatinib. b RT-PCR analysis showing downregulation of FOXA2 and SOX9 mRNA (Top 3 images) and immunoblot analysis of FOXA2 and β-actin (loading control) proteins (bottom 2 blots) in PCSC in response to afatinib treatment. c Transient suppression of EGFR in SP-SW1990 cells and its effects on CSC markers (FOXA2, SOX9, DCAMLK1, NANOG, and OCT4) and oncogenic signaling molecules (pERK) along with a reduction of total EGFR protein. d Immunoblot analysis of PCSC cells transiently knockdown for FOXA2 and its impact on SOX9 protein. e Immunoblot analysis showing a reduction in FOXA2 and SOX9 protein expression upon dose-dependent (IC20 and IC50 concentrations) treatments with afatinib alone in SPPC cells. f Immunofluorescence analysis of FOXA2 and SOX9 protein co-expression in afatinib-treated KPC tumoroids and mouse primary xenograft tumors. g Immunoblot analysis of FOXA2 and SOX9 protein expression in PC cells treated with afatinib/gemcitabine alone and combination therapies. h Schematic model of the hypothesized signaling axis by which afatinib inhibits the PCSC, scale bars= 5 μM and 2 μM.