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. 2017 Apr 24;127(6):2206–2221. doi: 10.1172/JCI91363

Figure 6. Knockdown of STRAP impairs erythroid differentiation, and overexpression of ITGB3BP is dispensable for granulomonocytic differentiation.

Figure 6

(A) Expression levels of STRAP determined using qRT-PCR in erythroid cells with STRAP knockdown. (B) Cell-cycle analysis of erythroid cells on day 11 of culture. (C) Images of erythroid cell pellets on day 14 of culture for visual determination of hemoglobinization (n = 6). (D) Percentage of late erythroid (CD71–CD235a+) cells on day 14 of culture. (E) Number of BFU-E obtained from hematopoietic CD34+ progenitors with STRAP knockdown after 14 days in methylcellulose (colony-forming cell assays). (F) Western blots showing the expression levels of the ITGB3BP protein in granulomonocytic cells with ITGB3BP overexpression. (G) Cell-cycle analysis of granulomonocytic cells on day 11 of culture. (H) Percentage of CD14+ cells in granulomonocytic cultures on day 20 of culture. (I) Percentage of CD15+ cells in granulomonocytic cultures on day 20 of culture. (J) Representative flow cytometric contour plots (from 6 independent experiments) showing expression of CD14 and CD15 on day 20 of culture. (K) Representative images of May-Grünwald-Giemsa–stained granulomonocytic cells on day 20 of culture (n = 6). The red arrows indicate eosinophils. Scale bars: 25 μm. Results in each bar graph in panels A, B, D, E, and G–I were obtained from 6 independent experiments. Data represent the mean ± SEM. P values in panels A, B, D, and E were calculated by repeated-measures 1-way ANOVA with Tukey’s post-hoc test. P values in panels H and I were calculated by a paired, 2-tailed t test. *P < 0.05, **P < 0.01, and ***P < 0.001.