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. 2023 Jun 27;30:16–26. doi: 10.1016/j.omto.2023.06.002

Figure 2.

Figure 2

Effects of RRSP-fusion proteins on Ras levels and viability

(A) Western Blot and quantification of the levels of β-actin and Ras proteins after incubation with different concentrations of Ec1-ETA-RRSP and RRSP-DT-Ec1 after 4 h of incubation in MCF7 cells (n = 3) A representative blot is shown. β-Actin (42 kDa) was used as a loading control. Ras proteins are detected at 21 kDa. (B) Western blot and quantification of Ras in MCF7 cell lysates after incubation with Ec1-ETA-RRSP and RRSP-DT-Ec1 at 100 nM or untreated control for different time points (n = 3). A representative blot is shown as in (A). (C) Cell viability of HCT116 cells after incubation with indicated concentrations of the proteins Ec1-ETA-RRSP, Ec1-ETA-RRSP∗, RRSP-DT-Ec1, and RRSP∗-DT-Ec1 (n = 3). RRSP∗ refers to a catalytically dead version of RRSP with the point mutation H451A. (D) Cell viability of MCF7 cells after incubation with different concentrations of the protein constructs (n = 3). (E) Cell viability in C5120 cells after incubation with different concentrations of the protein constructs (n = 3). Results in (C–E) are expressed as mean ± SEM.