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. Author manuscript; available in PMC: 2022 Apr 1.
Published in final edited form as: Cancer Res. 2019 May 22;79(14):3608–3621. doi: 10.1158/0008-5472.CAN-18-4055

Fig. 4. ONECUT2 is a negative regulator of CSC-associated properties.

Fig. 4.

(A) MDA231, MCF-7, and BT474 cells were transfected with siRNA against ONECUT2 (GeneSolution/GS including equal mixture of 4 preselected siRNAs, or individual siRNA #1 and #2), or a control siRNA, or with PBS. After 48 h, cells were collected for sphere formation assay. (B) ALDEFLUOR assay of MDA231 cells transfected with siRNA as indicated for 72 h. (C) RT-qPCR-determined RNA levels of indicated genes in various BC cells at 48 h following transfection with indicated siRNA. (D) Western blots showing the expression levels of indicated proteins in BC cells at 48 h following transfection of ONECUT2 siRNA-GS or control siRNA. (E) MDA231 cells were transfected with indicated siRNA and seeded at an equal number on day 0. DTX (10 nM) or DOXO (500 nM) was added on day 1, and replenished every 24 h. On day 3, cell viability (left) and cell number (right) were determined by MTS assay and cell counting, respectively, and compared to the PBS treatment group. (F) RT-qPCR of indicated genes in MDA231 cells at 72 h after transfection with indicated siRNA. *P<0.05, **P<0.01, ***P<0.001 compared to control siRNA or as indicated. Numbers below Western images indicate quantification after normalization to GAPDH with the first lane set as 1.