Figure 5: The C-terminal domain of LDHA recruits hCINAP to facilitate LDHA Y10 phosphorylation in a low-energy state. | Nature Communications

Figure 5: The C-terminal domain of LDHA recruits hCINAP to facilitate LDHA Y10 phosphorylation in a low-energy state.

From: Adenylate kinase hCINAP determines self-renewal of colorectal cancer stem cells by facilitating LDHA phosphorylation

Figure 5

(a) Schematic diagram of the structural domain organization within LDHA. (b,c) Co-IP analyses of the interaction between various regions of LDHA and hCINAP in SW480 cells. IgG served as the negative control. (d) Western blot analysis of LDHA Y10 phosphorylation level in CRCSCs transfected with indicated plasmids. (e) Immunofluorescence analysis of the effect of shRNA-depleted LDHA and LDHA/hCINAP rescue expression on the mesenchymal–epithelial transition in CRCSCs labelled by Vimentin (red), E-cadherin (green). The nucleus was counterstained with DAPI (blue). Scale bar, 100 μm. (f) Tumorsphere formation assay using cells in e. Scale bar, 100 μm. (g) Western blot analysis of LDHA Y10 phosphorylation in CRCSCs treated with different doses of 2-DG and glucose as indicated. (h) Co-IP assay for the interaction between endogenous hCINAP and LDHA protein in CRCSCs without glucose supply (de-glucose) or treated with 2-DG (50 mM) or oligomycin (1 μM). (i) The effects of 2-DG, glucose deprivation and oligomycin on LDHA Y10 phosphorylation in CRCSCs transfected with an empty vector or Flag-tagged hCINAP.

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