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. 2014 Jul 3;95(1):66–76. doi: 10.1016/j.ajhg.2014.06.003

Figure 4.

Figure 4

Validation of TREML4 as a Protein-Coding Gene

(A) RT-PCR results showing three bands for TREML4. Abbreviations are as follows: PB, peripheral blood; PMN, polymorphonuclear neutrophils; gDNA, genomic DNA; HaeIII, ladder with HaeIII-digested φx174 DNA (bands: from top, 1,353 bp, 1,072 bp, 872 bp, 603 bp); red star shows amplicon with uninterrupted CDS, alignment to TREML4 locus on chromosome 6 shown in lower panel.

(B) RNA blot results: left panel shows PMN RNA before probing, right panel shows bands for TREML4 after 25 hr exposure (black arrows) next to 2.4 kb marker band.

(C and D) Flow cytometry results from TREML4 construct with artificial CD8 leader sequence and FLAG tag showing detection on surface of HEK293 cells by αFLAG antibody (C) and anti-TREML4 antibody (D).

(E) Immunoblot results showing the larger protein from a construct with artificial leader sequence and smaller protein from construct with natural leader sequence. Mock transfection with vector only was used as control.