Figure 1. Silencing of PHB1 led to a decrease in cell proliferation and anchorage-dependent growth in HeLa cells.

(A, B) PHB1 mRNA and protein levels were efficiently reduced in cells expressing shPHB1-0, shPHB1-2 and shPHB1-3 using the constitutive lentiviral expression system pLL3.7 as was shown by qRT-PCR and Western Blots, respectively. (C) Upon lentiviral transduction stable integration is indicated by GFP expression. (D) With the expression of positively validated shRNAs targeting PHB1 mRNA the fraction of GFP expressing cells was reduced within ten days of transduction from a pool of transduced cells within, as shown by FACS analysis. (E) Transduced HeLa cells expressing control vector or PHB1 targeting shRNAs were seeded in soft agar. Only control cells grew under anchorage-independent conditions and formed colonies.