Supplementary Figure 11: LAIR1–SHP-1–CAMK1 axis supports human AML development.

(a) LAIR1^high primary AML cells have greater colony-forming ability difference in both first and second plating (samples B1 and B2), whereas no significant difference in colony-forming ability was detected between LAIR1^high and LAIR1^low cord blood mononuclear cells (sample A). Data from one experiment with n = 3 technical replicate samples are shown. The experiment was repeated 3 times with similar results. (b) Endogenous shp-1 expression was inhibited using shRNAs (201, 698, 786) in MV4-11 cells as determined by Q-PCR at 48 h after infection. Data are from a single experiment, representative of 3 independent experiments. (c) No clear association between shp-2 expression and AML patient survival was observed. Data were obtained from the TCGA AML database (n = 82 patient samples for high or n = 83 patient samples for low, p = 0.9451, log-rank test). (d) Endogenous camk1 expression was inhibited using an shRNA in MV4-11 cells as determined by Q-PCR at 48 h after infection. Data are from a single experiment, representative of 3 independent experiments. (e) LAIR1 expression is independent of the selected human AML stem cell phenotypic markers. The expression of LAIR1 and phenotypic markers (CD34/CD38/CD90) were analysed by flow cytometry in four AML clinical samples. (f) LAIR1 knockdown decreased colony-forming ability in all seven tested primary human AML cells as determined by CFU assays. Data from one experiment with n = 3 technical replicate samples are shown. The experiment was repeated 3 times with similar results. (g) The survival curves of mice receiving control or LAIR1-knockdown primary human patient AML cells (sample# 6). n = 9 mice; p < 0.0001, log-rank test. (h) Schematic summary of the novel signalling pathway mediated by the ITIM receptor LAIR1 in leukemia cells.