Fig. 5

Forced expression of SPI1 in H1-GATA2 ^−/− restores the generation of granulocytes upon OP9 co-culture. a, b Diagram of the strategy of SPI1 rescue experiments. SPI1 linked with a puromycin resistance gene by T2A sequence was controlled by a Dox-inducible promoter in lentiviral-based vectors for Dox-inducible expression of SPI1. The expression of SPI1 was not induced during later CFU assay. c Effects of enforced expression of SPI1 on generation of in CD34⁺ (left) and CD34⁺CD43⁺ (right) HPCs in H1-GATA2 ^−/−. Results are presented as mean + SEM of five independent experiments and normalized to H1 group. The data on CD34⁺ cells generation (left) were set as 1 for comparison. The data from five independent experiments were shown as box plot. Asterisks indicate statistical significance determined by t test: *p < 0.05, **p < 0.01 and ***p < 0.001. d Enforced expression of SPI1 in H1-GATA2 ^−/− regenerate G-CFUs. The error bars indicate mean + SEM of three independent experiments. e Morphology of CFU-G regenerated by SPI1 expression; bottom: cytospin of CFU-G. f, g FACS analysis of indicated markers in SPI1 regenerated CFU-M and CFU-G from H1-GATA2^−/−