Fig. 3
Downregulation of RHOB inhibits erythroid differentiation. (a-b) Downregulated RHOB expression in TF-1 cells (a) and MNCs (b) cultured in erythroid differentiation medium (TF-1 cells for 4 days, MNCs for 10 days) reduced the expression of erythroid markers (CD71 and CD235a). (c-d) Effect of siRHOB transfection on the percentage of mature RBCs (CD71−CD235+) from K562s with (c) or without (d) erythroid induction for 4 days. (e-h) Morphological and statistical analysis of erythroid cells with RHOB knockdown. Wright‒Giemsa staining revealed the erythroid morphology of TF-1 cells (e), MNCs (f) and K562 cells (g) after RHOB downregulation and erythroid induction (TF-1 and K562 cells for 4 days, MNCs for 10 days). (h) Cytospins of K562 cells cultured in complete medium were also stained with Wright‒Giemsa solutions. The nuclear area and cell area of more than 50 cells were measured from more than three random fields of view for each type of cell (scale bars: 50 μm). (i-k) Relative expression of erythroid marker genes in MNC-derived erythrocytes (i) and K562 cells with (j) or without (k) erythroid induction (K562 cells for 4 days, MNCs for 10 days). The data are presented as the means ± SDs.
