Journal: Science Advances

Article Title: Shifting IRES versus Cap-initiated translation during homeostatic stem cell differentiation and stress

doi: 10.1126/sciadv.adz7896

Figure Lengend Snippet: ( A ) Representative gating strategy for sorting low, mid, and high IRES/Cap SLAM-LKS. ( B and C ) Colony formation (B) and size (C, mm 2 ) of Translator SLAM-LKS sorted on the basis of IRES/Cap ( n = 6). ( D and E ) Colony formation (D) and size (E, mm 2 ) of Translator GMP sorted on the basis of IRES/Cap ( n = 9). ( F ) Total protein synthesis rates of Translator GMP based on IRES/Cap measured by AF647-OPP MFI ( n = 3). ( G ) Colony formation of EMCV-IRES reporter-transduced CD34+CD90+EPCR+CD45RA− human cord blood sorted on the basis of IRES/Cap ( n = 3). ( H and I ) Colony formation of SLAM-LKS (H) and GMPs (I) based on RUNX1-IRES/Cap sorted from transplant recipients ( n = 3 and 2, respectively). ( J ) Primary competitive transplant schema. Twenty CD45.2 SLAM-LKS sorted on the basis of IRES/Cap versus 200,000 CD45.1 whole BM cells in lethally irradiated CD45.1 recipients. Peripheral blood chimerism analyzed every four weeks through week 16. Created in BioRender. Li, D. (2026) https://BioRender.com/d7k2abt . ( K ) Total chimerism Translator SLAM-LKS based on IRES/Cap in primary recipients evaluated through week 16 ( n = 12). ( L ) Secondary competitive transplant schema. One million total BM cells were collected from primary recipients (16 weeks posttransplantation) and transplanted to lethally irradiated CD45.1 recipients. Created in BioRender. Li, D. (2026) https://BioRender.com/d7k2abt . ( M ) Total chimerism Translator SLAM-LKS based on IRES/Cap in secondary recipients through week 16 ( n = 12). ( N and O ) Representative image of the culture of a single low (N) or high (O) IRES/Cap SLAM-LKS after 5 days of culture. ( P and Q ) Pie charts depicting the percentage of wells with megakaryocytes (MK) after 5 days of culturing a single low (P) or high (Q) IRES/Cap Translator SLAM-LKS ( n = 20). Data show individual replicates and means ± SEM. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001. Significance was assessed using a two-way ANOVA with Dunnett’s one-way ANOVA (B to I) or an ANOVA with Tukey’s multiple comparisons test (K and M).

Article Snippet: WT CD45.2 C57BL/6J (strain #000664), CD45.1 (B6.SJL- Ptprc a Pepc b /BoyJ; strain #002014), KH2/iCas9 [B6;129S4- Gt(ROSA)26Sor tm1(rtTA*M2)Jae Col1a1 tm1(tetO-cas9)Sho /J; strain #029415], Rpl24 Bst (C57BLKS-Rpl24 Bst /J; strain #000516), and NSG mice (NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ; strain #005557) were purchased from the Jackson Laboratory.

Techniques: Irradiation

Journal: Science Advances

Article Title: Shifting IRES versus Cap-initiated translation during homeostatic stem cell differentiation and stress

doi: 10.1126/sciadv.adz7896

Figure Lengend Snippet: ( A ) Representative gating strategy for sorting low, mid, and high IRES/Cap SLAM-LKS. ( B and C ) Colony formation (B) and size (C, mm 2 ) of Translator SLAM-LKS sorted on the basis of IRES/Cap ( n = 6). ( D and E ) Colony formation (D) and size (E, mm 2 ) of Translator GMP sorted on the basis of IRES/Cap ( n = 9). ( F ) Total protein synthesis rates of Translator GMP based on IRES/Cap measured by AF647-OPP MFI ( n = 3). ( G ) Colony formation of EMCV-IRES reporter-transduced CD34+CD90+EPCR+CD45RA− human cord blood sorted on the basis of IRES/Cap ( n = 3). ( H and I ) Colony formation of SLAM-LKS (H) and GMPs (I) based on RUNX1-IRES/Cap sorted from transplant recipients ( n = 3 and 2, respectively). ( J ) Primary competitive transplant schema. Twenty CD45.2 SLAM-LKS sorted on the basis of IRES/Cap versus 200,000 CD45.1 whole BM cells in lethally irradiated CD45.1 recipients. Peripheral blood chimerism analyzed every four weeks through week 16. Created in BioRender. Li, D. (2026) https://BioRender.com/d7k2abt . ( K ) Total chimerism Translator SLAM-LKS based on IRES/Cap in primary recipients evaluated through week 16 ( n = 12). ( L ) Secondary competitive transplant schema. One million total BM cells were collected from primary recipients (16 weeks posttransplantation) and transplanted to lethally irradiated CD45.1 recipients. Created in BioRender. Li, D. (2026) https://BioRender.com/d7k2abt . ( M ) Total chimerism Translator SLAM-LKS based on IRES/Cap in secondary recipients through week 16 ( n = 12). ( N and O ) Representative image of the culture of a single low (N) or high (O) IRES/Cap SLAM-LKS after 5 days of culture. ( P and Q ) Pie charts depicting the percentage of wells with megakaryocytes (MK) after 5 days of culturing a single low (P) or high (Q) IRES/Cap Translator SLAM-LKS ( n = 20). Data show individual replicates and means ± SEM. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001. Significance was assessed using a two-way ANOVA with Dunnett’s one-way ANOVA (B to I) or an ANOVA with Tukey’s multiple comparisons test (K and M).

Article Snippet: Heterozygous transgene positive founders were bred with WT CD45.2 C57BL/6J mice purchased from the Jackson Laboratory.

Techniques: Irradiation