Skip to main content
. 2019 Jan 24;9(3):811–828. doi: 10.7150/thno.29271

Figure 1.

Figure 1

RACK1 promotes self-renewal and chemoresistance of human liver CSCs and maintains murine ESC function. (A-N) 96 h after HuH7 cells were infected with lentivirus expressing non-targeting control (NC) shRNA or RACK1 shRNA (A-G), or after HuH7 single clone stably expressing FLAG-RACK1 and the mock control were generated (H-N), cells were subjected to the following assays: (A,H) Immunoblotting (IB) analysis of RACK1 expression in total cells. (B,I) Flow cytometric analysis of CD13 and CD133 expression in total cells. (C,J) Immunoblotting analysis of RACK1 expression in sorted CD13+ and CD13- subpopulations. (D,K) Sphere formation assays of sorted CD13+ subpopulation. mean±s.d. (n=3); *P<0.05, **P<0.01. (E,L) In vivo tumorigenicity experiments of sorted CD13+ subpopulation (5000 cells/site, 7 weeks, n=6). (F,M) Etoposide (Etop, 100 μM, 48 h)- or sorafenib (Sora, 50 μM, 24 h)-induced apoptosis of CD13+ subpopulation. mean±s.d. (n=3); Ctrl, control. (G,N) qRT-PCR analysis of sorted CD13+ subpopulation for the expression of the indicated drug-resistant relative genes. mean±s.d. (n=3). (O-Q) 96 h after murine ESCs were infected with lentivirus expressing non-targeting control shRNA or RACK1 shRNAs, cells were subjected to immunoblotting analysis for RACK1 expression (O), colony formation assays (mean±s.d., n=3; scale bar: 1 cm) (P), and alkaline phosphatase (AP) activity assays (mean±s.d., n=3) (Q).