Extended Data Fig. 7: Related to conditional HK2 dependence varies with cell-intrinsic factors.
(a) HK signal normalized by COXIV signal in IP:HA versus WC from the same respective cells. nd, not detected. (b) Immunoblots for expression of HK1 and HK2 (c) in dounce homogenates (DH) and corresponding cytosolic (Cyt) and organellar (Org) fractions isolated from NOMO1 cells. S6K served as cytosolic control marker. COXIV served as a mitochondrial control marker. L.B., low brightness; H.B., high brightness. (c) HK signal normalized by COXIV signal in Org versus DH from the same respective cells. Comparisons of normalized signal between HK1 and HK2 are derived using the same experimental samples run across different blots and processed in parallel. (d) Relative growth of indicated cell lines transduced with sgHK2 versus sgAAVS1 (mean ± s.d., n = 3 biologically independent samples). Two-tailed Welch’s t-test comparing the respective mean ± s.d. (bar) versus mean ± s.d. (control cells) between bars. (e) Relative growth rates of indicated cell lines transduced with sgHK2 versus sgAAVS1 (mean ± s.d., n = 3 biologically independent samples). Two-tailed Welch’s t-test comparing the respective mean ± s.d. (bar) versus mean ± s.d. (control cells) between bars. (f) Distribution of mRNA transcript levels for indicated monocarboxylate transporter (MCT)-encoding SLC16 isoforms across human cancer lines from cataloged RNA-Seq data23. Displayed percentage, cell lines with log2(TPM + 1) greater than 1. Bolded line, median. TPM, transcripts per million. (g) mRNA transcript levels for SLC16A7 (top) and SLC16A8 (bottom) across the indicated cell lines from analyzed RNA-seq data80. SLC16A7 encodes MCT2. SLC16A8 encodes MCT3. nTPM, normalized TPM. (h) Human cancer lines ranked by SLC16A3 mRNA transcript levels from cataloged RNA-Seq data23. Labeled points indicate cell lines that exhibited conditional HK2 dependence in this study.
