Fig. 1
CXCR4 expression in AML cell lines. (A) RT-PCR of CXCR4, ACKR3, and β actin in three human AML cell lines U937, THP-1, and AML-193. Data representative of 3 separate analyses. (B) Flow cytometry results of the three AML cell lines stained with either anti-CXCR4 clone 12G5 or control isotype antibody. * = P ≤ 0.05 by Student’s t-test. n = 2–3 independent analyses. Values are mean ± SEM. (C) U937 or (D) THP-1 cell chemotaxis from the top chamber of a transwell filter towards the lower chamber containing the indicated concentration of CXCL12-WT chemokine. All values under significance bar represent a significant increase by one-way ANOVA with Dunnett’s multiple comparison test to 0 nM vehicle treated. (E) Total number of U937 or (F) THP-1 migrated from the top chamber of a transwell chamber towards the lower chamber containing the indicated concentration of CXCL12-WT, CXCL12-LM, or CXCL12-LD. Migrating cells in panels C-F were enumerated after a 2 h incubation using flow cytometry. Values represent mean ± SEM, n = 3–4 independent biological replicates. Red asterisks or numbers indicate adjusted p value by two-way ANOVA with Tukey’s multiple comparisons test to that respective variant’s 0 nM (e.g. CXCL12-WT 30 nM to CXCL12-WT 0 nM) while blue indicates significance compared to the same dose of a different variant (e.g. CXCL12-LD 30 nM to CXCL12-WT 30 nM).
