Extended Data Fig. 6: Effect of lactate accumulation on cyclin B1, securin, and mitotic exit.
From: Lactate regulates cell cycle by remodelling the anaphase promoting complex
(a) Model for putative role of chronic lactate elevation on APC/C dependent processes in proliferative cells. (b) Intracellular lactate concentration in asynchronous HeLa S3 (n = 6), DLD1 (n = 4), and HCT116 (n = 5) cells. n indicates cell replicate number. (c) Treatment of asynchronous human proliferative cells with elevated lactate at 15 and 25 mM for 4 h lowers levels of securin and cyclin B1. Representative blot from three experiments shown. (d) mRNA transcript abundance of human FOXK1 (n = 3) and FOXK2 (n = 4) in HCT116 cells following engineered overexpression (oe). n designates cell replicates. (e) Effect of forced overexpression on abundance of cyclin B1 and securin in asynchronous HCT116 cells. Representative blot from two experiments shown. (f) Treatment of asynchronous human proliferative cells with elevated glucose for 4 h has no effect on levels of securin and cyclin B1. Representative blot from two experiments shown. (g) Treatment of asynchronous human proliferative APC4 K772/798R cells with 25 mM lactate for 4 h has no effect on securin or cyclin B1 abundance. Representative blot from three experiments shown. (h) Lactate mediated depletion of securin and cyclin B1 is lost upon treatment with the proteasome inhibitor MG132. Representative blot from three experiments shown. (i) Treatment of asynchronous HCT116 cells with 15 mM lactate for 4 h lowers levels of securin and cyclin B1 in the presence of the LDH inhibitor GSK2837808A. Representative blot from two experiments shown. (j) Effect of treatment of asynchronous human proliferative cells with 1 µM oligomycin A for 24 h on abundance of securin and cyclin B1 in WT and APC4 K772/798R cells. Representative blot from three experiments shown. (k) Effect of treatment of asynchronous human proliferative cells with 5 mM αCHCA for 24 h on abundance of securin and cyclin B1 in WT and APC4 K772/798R cells. Representative blot from three experiments shown. (two-tailed Student’s t-test for pairwise comparisons in d). Data are mean ± s.e.m.
