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. 2015 Mar 27;10(3):e0123241. doi: 10.1371/journal.pone.0123241

Fig 4. Rescue of SPP1 RNAi growth defect by expression of recoded SPPI.

Fig 4

(a) Expression of recoded HA-tagged SPP1 detected by Western blot. Cell lysates from RNAi cell line (-), the RNAi cell line expressing SPP1 from the recoded gene (SPP1 R), or the RNAi cell line expressing inactive SPP1 from a recodedgene (SPP1 R,I) were probed with anti-HA antibody (Roche). Detection of EF-1α was used as a loading control. (b, c and e) Parasite growth was measured in cell lines after inducing SPP1 RNAi with tetracycline (open squares) or without treatment (closed squares) in the SPP1 RNAi cell line expressing SPP1 from: the recoded gene SPP1 R (b), the parental SPP1 RNAi cell line (c), or the RNAi cell line expressing inactive SPP1 from a recoded gene, SPP1 R,I (e). (d) Quantitative PCR showing relative quantification (RQ) of endogenous SPP1 transcript in the RNAi cell line expressing active SPP1 from the SPP1 R gene either without induction (control, black bars) or after induction by tetracycline (+ Tet, white bars).