Supplementary Figure 4: Lair1, a representative ITIM receptor, is essential for the growth of human leukemia cell lines.

(a) Expression of certain human ITIM receptor mRNAs negatively correlates with the overall survival of AML patients. A total of 58 ITIM receptors were selected based on the criteria that (1) they are plasma membrane receptors and (2) they use ITIM as the main signalling motifs. Data were obtained from the TCGA AML database, and analysed without normalization (condition 1) or normalized to GADPH expression (condition 2), Affymetrix housekeeping gene expression (condition 3), or total mRNA (condition 4). More information about data analysis can be found in Methods. (b) Effects of inhibition of expression of indicated ITIM receptors using shRNAs as determined by real-time RT-PCR. Data are from a single experiment, representative of 3 independent experiments. (c) In silico analysis of the correlation between human lair1 mRNA expression and the overall survival of AML patients younger than 65 years old. Data were obtained from the TCGA AML database (n = 52 patient samples for each groups, p = 0.0271, log-rank test). (d) An in silico analysis of human lair1 mRNA expression in 43 human AML samples as described previously¹. (e) SP-D is not highly expressed in the bone marrow environment compared with mRNA expression in lung epithelial cells, as determined by real-time RT-PCR. Data are from a single experiment, representative of 3 independent experiments. (f) Schematic summary of the LAIR1 chimeric receptor signalling reporter cells system. (g) In the LAIR1 chimeric receptor signalling reporter cells system, flow cytometry analysis demonstrated that, while the immobilized collagen 1 (1 μg ml⁻¹) or anti-LAIR1 antibody induced LAIR1 activation (as shown by increased GFP induction), the immobilized or soluble SP-D (1 μg ml⁻¹) was unable to do so (the upper panels are for control cells, and the lower panels are for the hLAIR1 reporter cells).