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. 2010 Oct 1;21(19):3340–3351. doi: 10.1091/mbc.E10-01-0040

Figure 5.

Figure 5.

Stat5b silencing is sufficient to down-regulate the expression of erythroid markers and to impair erythroid differentiation of CD34+ HSCs. (A) Western blot analysis showing the down-regulation of Stat5b obtained in Hel cells transfected with anti-Stat5b siRNAs (lane 2) compared with cells transfected with a nontargeting control siRNA (lane 1); cells were lysed 48 h after transfection. (B) Real-time PCR performed on Hel cells transfected with control siRNA (□) or with anti-Stat5b siRNAs (▩) demonstrating the efficacy of Stat5b silencing and the down-regulation of erythroid markers; ACTB was used as control housekeeping gene (error bars, SEM calculated on a set of three experiments; *p < 0.05; **p < 0.01). RNAs were extracted 48 h after transfection with siRNAs. (C) Western blot analysis showing the down-regulation of Stat5b obtained in CD34+ HSCs transfected with anti-Stat5b siRNAs (lane 2) compared with cells transfected with a nontargeting control siRNA (lane 1); cells were lysed 7 d after transfection. (D) Real-time PCR performed on CD34+ HSCs transfected with control siRNA (□) or with anti-Stat5b siRNAs (■) demonstrating the efficacy of Stat5b silencing and the down-regulation of erythroid markers; ACTB was used as control housekeeping gene (error bars, SEM calculated on a set of three experiments; *p < 0.05; **p < 0.01). RNAs were extracted 96 h after transfection with siRNAs. (E) Clonogenic assay performed on CD34+ HSCs transfected with a nontargeting control siRNA (left) or with anti-Stat5b siRNAs (right); SEM calculated on three independent experiments; *p < 0.05.