Figure 5.
RNA editing capability of ADAR2 is essential for the suppression of leukemogenesis in an AE9a mouse model. (A) Experimental strategy for AE9a BM transplantation mouse model. (B) Bar chart showing the relative expression of ADAR2 transcripts in BM cells from moribund mice after the first transplantation, and in the second transplantation injected with different numbers of AE9a cells (50 000, 100 000, and 200 000). The relative expression was calculated as described in supplemental Materials and Methods. Data are presented as the mean ± SD of technical triplicates from a representative experiment. (C) Western blot analysis of ADAR2 protein expression in AE9a BM cells stably overexpressing ADAR2 or the MSCV-IRES-tdTOMATO EV. (D) Dot plot representing counts of white blood cells (WBCs), red blood cells (RBCs), and platelets in the PB from recipients at 28 days after transplantation of AE9a AML cells stably overexpressing AE9a or the MSCV-IRES-tdTOMATO EV. n = 5 in each group. (E) Kaplan-Meier survival curve of recipients who underwent transplantation with 50 000 AE9a AML cells stably overexpressing ADAR2 or MSCV-IRES-tdTOMATO EV. n = 16 in each group. Statistical analysis was performed using log-rank (Mantel-Cox) test. (F) Western blot analysis of ADAR2 or ADAR2 DeAD protein expression in BM cells from the same recipients the indicated groups of cells described in panel E. Actin was used as a loading control. (G) Dot plot representing counts of WBCs, RBCs, and platelets in the PB from recipients at 45 days after transplantation of AE9a AML cells stably overexpressing of ADAR2 (n = 6), ADAR2 DeAD (n = 5), or MSCV-IRES-tdTOMATO EV (n = 6). (H) Kaplan-Meier survival curve of recipients who underwent transplantation with 50 000 AE9a AML cells stably overexpressing ADAR2 (n = 9), ADAR2DeAD (n = 7), or MSCV-IRES-tdTOMATO EV (n = 9). Statistical analysis was performed using log-rank (Mantel-Cox) test. FL, full-length; IRES, internal ribosome entry site. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001 using two-tailed Student t test.

RNA editing capability of ADAR2 is essential for the suppression of leukemogenesis in an AE9a mouse model. (A) Experimental strategy for AE9a BM transplantation mouse model. (B) Bar chart showing the relative expression of ADAR2 transcripts in BM cells from moribund mice after the first transplantation, and in the second transplantation injected with different numbers of AE9a cells (50 000, 100 000, and 200 000). The relative expression was calculated as described in supplemental Materials and Methods. Data are presented as the mean ± SD of technical triplicates from a representative experiment. (C) Western blot analysis of ADAR2 protein expression in AE9a BM cells stably overexpressing ADAR2 or the MSCV-IRES-tdTOMATO EV. (D) Dot plot representing counts of white blood cells (WBCs), red blood cells (RBCs), and platelets in the PB from recipients at 28 days after transplantation of AE9a AML cells stably overexpressing AE9a or the MSCV-IRES-tdTOMATO EV. n = 5 in each group. (E) Kaplan-Meier survival curve of recipients who underwent transplantation with 50 000 AE9a AML cells stably overexpressing ADAR2 or MSCV-IRES-tdTOMATO EV. n = 16 in each group. Statistical analysis was performed using log-rank (Mantel-Cox) test. (F) Western blot analysis of ADAR2 or ADAR2 DeAD protein expression in BM cells from the same recipients the indicated groups of cells described in panel E. Actin was used as a loading control. (G) Dot plot representing counts of WBCs, RBCs, and platelets in the PB from recipients at 45 days after transplantation of AE9a AML cells stably overexpressing of ADAR2 (n = 6), ADAR2 DeAD (n = 5), or MSCV-IRES-tdTOMATO EV (n = 6). (H) Kaplan-Meier survival curve of recipients who underwent transplantation with 50 000 AE9a AML cells stably overexpressing ADAR2 (n = 9), ADAR2DeAD (n = 7), or MSCV-IRES-tdTOMATO EV (n = 9). Statistical analysis was performed using log-rank (Mantel-Cox) test. FL, full-length; IRES, internal ribosome entry site. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001 using two-tailed Student t test.

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