Fig. 2

Knockdown of ASF1B in human primary erythroid cells induces γ-globin expression. a Expression analysis by RT-qPCR of ASF1B, BCL11A, and KLF1 (last two taken from Fig. 1b) on shRNA lentivirus-transduced HEP cultures. Expression levels are normalized setting expression levels of mock knockdown controls (Scr) at 100. Mean and standard deviation are depicted. Shadowed box at 100 corresponds to the expression levels in mock knockdown controls. T test p values are indicated. b The expression of BCL11A and KLF1 is not significantly affected in ASF1B knockdown HEPs as analyzed by RT-qPCR. Expression levels are normalized setting expression levels of mock knockdown controls (Scr) at 100. Mean and standard deviation are depicted. Shadowed box at 100 corresponds to the expression levels in mock knockdown controls. T test p values are indicated. c HBB and HBA expression analysis by RT-qPCR. Knockdown of either KLF1 or BCL11A has an effect on HBB expression, whereas ASF1B knockdown does not, while overall, HBA expression is not affected. Expression levels are normalized setting expression levels of mock knockdown controls (Scr) at 100. Mean and standard deviation are depicted. Shadowed box at 100 corresponds to the expression levels in mock knockdown controls. T test p values are indicated. d RT-qPCR of HBG expression and HPLC analysis of hemoglobin composition show the increase in HbF after knockdown of ASF1B in HEPs. Values are normalized to those measured in mock knockdown controls (Scr). Mean and standard deviation are depicted. Shadowed box at 100 corresponds to the expression levels in mock knockdown controls. T test p values are indicated