Journal: Cell reports
Article Title: RSPO2 inhibits BMP signaling to promote self-renewal in acute myeloid leukemia.
doi: 10.1016/j.celrep.2021.109559
Figure Lengend Snippet: Figure 3. RSPO2 is required for proliferation of AML cells (A) Cumulative cell growth curve depicted as the total cell count of THP-1 shRNA cell lines with Dox treatment. Cell number was counted at the indicated time points with automated cell counter and normalized to the initial value. n = 3 per group. (B) Cell-cycle FACS analysis using BrdU/PI double staining of THP-1 shRNA cell lines. Cells were treated with Dox for 4 days and incubated with 10 mM BrdU for 1 h before FACS analysis. n = 3 per group. PI, propidium iodide. (C) Experimental scheme of colony formation cell (CFC) assay with Dox-inducible shRNA THP-1 cell lines. (D) Representative images of CFC assay of THP-1 colonies. Scale bar, 200 mM. (E) Quantification of THP-1 CFC assay. n = 3 per group. (F) Representative images of CFC assay of MOLM14 colonies. Scale bar, 200 mM. (G) Quantification of MOLM14 CFC assay. n = 3 per group. (H) Quantification of FACS analysis for active caspase-3+ cells in THP-1 clones with Dox treatment for 3 days. Cells were fixed by paraformaldehyde (PFA) and permeablized with MeOH before Ab staining. n = 3 per group. (I) AraC IC50 of THP-1 clones upon Dox treatment. TetOn shRSPO2 cells were pretreated with Dox for 2 days. IC50 was determined after 3 days incubation with increasing amount of AraC in the presence of Dox as indicated. n = 3 per group. Results are presented as the mean ± SD. ns, not significant; *p < 0.05, **p < 0.01,***p < 0.001 from unpaired t test for experiments with two groups or one-way ANOVA for experiments with more than two groups. See also Figure S3.
Article Snippet: Cytarabine/AraC IC50 measurement and proliferation assay Equal numbers of AML cells were treated with increasing concentrations of cytarabine (Selleck Chemicals) for 72 hours.
Techniques: Cell Counting, shRNA, Double Staining, Incubation, Clone Assay, Staining
