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. 2020 Mar 10;28(5):1287–1298. doi: 10.1016/j.ymthe.2020.03.002

Figure 3.

Figure 3

circSKA3 Interacts with Tks5 and Integrin β1

(A) Photographs showed that expressing circSKA3 enhanced MCF-7 cell invadopodium formation. (B) Left: cell lysates from circSKA3- or vector-transfected cells were subjected to immunoprecipitation with antibodies against proteins as shown. Anti-Tks5, integrin β1 (Itgb1), ERK, MMP2, MMP11, and Cdc42 antibodies pulled down high levels of circSKA3 in circSKA3-overexpressing cells compared with the control. Right: cell lysates were subjected to immunoprecipitation followed by real-time PCR with primers specific for linear SKA3 mRNA or circSKA3. Antibodies against Tks5, Itgb1, EGF, MMP2, MMP11, and F-actin pulled down circSKA3, but not the linear SKA3 mRNA. ∗∗p < 0.01. Error bars, SD (n = 4). (C) Left: real-time PCR showed expression of circSKA3 in the inputs. Middle: PCR showed that circSKA3 probe pulled down higher levels of circSKA3 in the circSKA3-transfected cells relative to controls. Right: in the circSKA3-transfected cell lysate, circSKA3 probe pulled down more Itgb1 and Tks5 relative to controls. ∗∗p < 0.01. Error bars, SD (n = 4). (D) Left: real-time PCR showed expression of circSKA3 in the inputs. Right: PCR showed that circSKA3 pulled down lower levels of circSKA3 in the circSKA3 siRNA-transfected cells relative to controls. ∗∗p < 0.01. Error bars, SD (n = 4). (E) circSKA3 probe precipitated Itgb1 and Tks5, which were decreased by circSKA3 siRNAs. (F) Immunoprecipitation assay showed that anti-Tks5 and -Itgb1 antibodies pulled down higher levels of circSKA3 in controls compared with the siRNA-transfected cells. ∗∗p < 0.01. Error bars, SD (n = 4).