Fig. 7: TLR8 inhibition improves the number and differentiation of erythroid cells generated by cell lines and primary cells from healthy donors and DBA patients.
From: Erythroid-intrinsic activation of TLR8 impairs erythropoiesis in inherited anemia
a Representative immunoblotting showing RPS19 levels in WT and RPS19+/− HuDEP2 cells and the quantitation (n = 3 biologically independent experiments). b Schematic illustrating the experimental design for analysis of RPS19+/− HuDEP2 cell lines. Lower panel shows representative levels of CD235a expression in WT and RPS19+/− HuDEP2 cells after treatment with 10 µM CUCPT8m for 4 days (n = 4 biologically independent experiments). c Quantification of MFI of CD235a expression shown in (b) (n = 4 biologically independent experiments). d Total cell numbers of WT and RPS19+/− HuDEP2 cells treated with/without 10 μM CUCPT8m. 2 × 105 cells are seeded accompanied with 10 μM CUCPT8m treatment for 4 days. Total cell number is then counted (n = 4 biologically independent experiments). e Representative immunoblotting showing levels of p-STAT5 and t-STAT5 in WT and RPS19+/− HuDEP2 cells treated with 3 U/ml EPO alone or in combination with 10 μM CUCPT8m for 2 h and f quantification of relative protein expression (p-STAT5/t-STAT5; n = 3 biologically independent experiments). g Schematic illustrating erythroid differentiation and analysis of CD34+ cells from RP-mutant DBA patient. h Flow cytometry results showing erythroid differentiation (days 8 and 11) of BM-derived CD34+ cells from a RPS19-mutant DBA patient after consecutive treatment with 10 μM CUCPT8m. i Cumulative proliferation curve of BM-derived CD34+ cells from two RPL5-mutant DBA patients treated with/without TLR8 inhibitors. Data are mean ± SEM; statistical significance was determined using the unpaired two-tailed Student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001, ns = not significant. Source data are provided as a Source Data file.
