1. Droplet Digital PCR for Absolute Quantification of EML4-ALK Gene Rearrangement in Lung Adenocarcinoma 
Qiushi Wang, Xin Yang, Yong He, Qiang Ma, Li Lin, Ping Fu, Hualiang Xiao 
The Journal of Molecular Diagnostics 
Volume 17, Issue 5, Pages (September 2015)
DOI: /j.jmoldx
Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

2. Figure 1
ALK gene rearrangement results for 103 clinical formalin-fixed, paraffin-embedded samples detected by fluorescent in situ hybridization (FISH) and Droplet Digital PCR (ddPCR). For easy identification, some of the negative samples (colored gray) were truncated. A: ALK status detected by FISH (15% was used as the cutoff point for ALK positivity). B: Normalized variant 1 (v1 nor) results of EML4-ALK. C: Normalized variant 2 (v2 nor) results of EML4-ALK. D: Normalized variant 3 (v3 nor) results of EML4-ALK; 14 ALK-positive samples detected by FISH (black, red, green, and blue; A) were also assessed via ddPCR for EML4-ALK copy number alterations. The results are highlighted by the red (B), green (C), and blue (D) colors. The specimen numbered (30% ALK break-apart signals by FISH test, colored black; A) failed the ddPCR test and finally identified as KIF5B-ALK gene rearrangement by PCR-based direct sequencing.
The Journal of Molecular Diagnostics  , DOI: ( /j.jmoldx )
Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

3. Figure 2
Fluorescent in situ hybridization (FISH) and Droplet Digital PCR (ddPCR) tests reveal low copy numbers of EML4-ALK fusion in a patient successfully treated with crizotinib. A: FISH test revealed 10% tumor cells harbored break-apart ALK signals, which failed the positive cutoff point of 15% (arrows indicate break-apart ALK signals). B: ddPCR test revealed low copy numbers of EML4-ALK translocation (normalized variant 3 =  ). Nor., normalized; V, version.
The Journal of Molecular Diagnostics  , DOI: ( /j.jmoldx )
Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

4. Figure 3
Chest computed tomography (CT) revealing a decrease in the primary tumor in a patient with 10% (EML4-ALK/GAPDH =  , variant 3) ALK rearranged tumor cells after 7 months of crizotinib treatment (partial response). A: CT scan before crizotinib treatment. B: CT scan after 7 months of crizotinib treatment. Arrows indicate decreased tumor and nonreduced pleural effusion.
The Journal of Molecular Diagnostics  , DOI: ( /j.jmoldx )
Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

5. Supplemental Figure S1
Raw data of Droplet Digital PCR (ddPCR) for false-positive control. Eight samples of reactive lymph nodes were used as the false-positive control of the ddPCR test and finally revealed negligible positive signals.
The Journal of Molecular Diagnostics  , DOI: ( /j.jmoldx )
Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions