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Fig. 3 | Genome Medicine

Fig. 3

From: Functional fingerprinting of human mesenchymal stem cells using high-throughput RNAi screening

Fig. 3

Validation of screening hits identified multiple kinases regulating MSC viability. a Cell viability was determined 72 h after siRNA reverse transfection (ATP level measured by luminescence) and the 19 genes which revealed the strongest phenotype are depicted. relative light units (RLU) were normalized to the negative control Rluc siRNA (red line). Data are presented as mean ± standard deviation (s.d.) of three screens. b Representative microscopy images of selected candidates revealed differences in morphology. Cells were reverse transfected with the corresponding siRNA and stained for microtubules (FITC), actin filaments (Alexa Fluor 547) and DNA (Hoechst). c Single siRNAs from selected candidates show good knockdown phenotype correlation. Single siRNAs and corresponding siRNA pools were analyzed for relative cell viability (CellTiterGlo) and knockdown efficiency (quantitative PCR) 72 h after reverse transfection. Relative viability compared to controls ± s.d. Significance of ATP level changes were calculated using unpaired two-tailed student’s T-test; n ≥ 3; *p ≤ 0.05; **p ≤ 0.01

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