1. Relative to 18S rRNA
Supplementary Figure S1. H2228 and H3122 cells express HK1 and HK2 mRNAs.
The qPCR was performed as described in Figure 5 and the results were presented as values relative to 18S rRNA.

2. 5’-GCGTGCGCACGT-3’
5’-GAAAGCGCTTTT-3’
5’-RCGTGXXCACGY-3’
HRE in the HK2 promoter
HRE mutant of the HK2 promoter
Canonical HRE
Supplementary Figure S2. The wild type and mutant versions of the HRE in the human HK2 gene promoter are aligned and compared with the sequence of the canonical HRE. The functional HRE is composed of a pair of contiguous sites 5′-RCGTG-3′ (underlined). Y: a pyrimidine; R: a purine; XX: 1-8 of any residue. The numbering is based on the human HK2 gene promoter.

3. VHL
H2228
HIF1a
b-tubulin
H3122
HIF2a
CoCl
Ceritinib
Supplementary Figure S3. EML4-ALK selectively upregulates HIF1a, but not HIF2a, under normoxic conditions. H2228 and H3122 were treated with 100 μM CoCl2 or 200 nM Ceritinib for 2 hours before immunoblotting analysis with indicated antibodies.

4. **
LDHA mRNA
GLUT1 mRNA
EML4-ALK
Vector
LDHA
GLUT1
HK2
b-tubulin
ALK
A549
mRNA expression level A B C D
Supplementary Figure S4. EML4-ALK upregulates GLUT1 and LDHA mRNA despite no effect on their proteins. The qPCR analysis was performed to examine the effects of EML4-ALK knockdown on levels of GLUT1 (A) and LDHA (B) mRNA in H2228 and H3122 cells. The lower panel shows the effects of EML4-ALK stable overexpression in A549 cells on GLUT1 and LDHA mRNAs analyzed by qPCR (C) and their proteins by immunoblotting (D). The qPCR probes for GLUT1 (Hs ) and LDHA (Hs ) were obtained from Life Technologies. The qPCR results were normalized to 18S rRNA and presented as values relative to the corresponding control cells (defined as 1 fold).

5. immunoblotting analysis of 4E-BP1 phosphorylation.
Glucose g/l
p-4E-BP1T37/46
b-tubulin
H2228
H3122
Supplementary Figure S5. Glucose concentrations do not affect 4E-BP1 phosphorylation.
H2228 and H3122 cells were cultured with indicated concentrations of glucose for 24 hours before
immunoblotting analysis of 4E-BP1 phosphorylation.