Fig. 3: Media profiling reveals that asparagine rescues inhibition of respiration. | Nature Cancer

Fig. 3: Media profiling reveals that asparagine rescues inhibition of respiration.

From: Differential integrated stress response and asparagine production drive symbiosis and therapy resistance of pancreatic adenocarcinoma cells

Fig. 3

a, Differentially consumed/released metabolites present in the media from group 1 and group 2 clones (n = 3 replicates per cell line) after 48 h of culture. b, Relative abundance of NEAAs present in conditioned medium at higher levels than in basal medium (n = 3 replicates per cell line). c, Doubling time of clone N treated with 1 nM oligomycin in the presence or absence of medium containing a 100 µM cocktail of all NEAAs (n = 3 samples per condition). d, Doubling times of clone N treated with 1 nM oligomycin in the presence or absence of individual NEAAs (n = 3 samples per condition). e, Doubling times of clone N treated with 1 nM oligomycin in the presence or absence of 100 µM or 20 mM aspartate (Asp; n = 3 samples per condition). f, Doubling times of clonal line N treated with 1 nM oligomycin, 25 µM phenformin or 25 nM IACS-10759 ± asparagine (Asn). g, Clonal line N was encoded with a fluorescent label and plated in direct coculture with unlabeled clones transfected with siRNA targeting Asns or non-targeted (NT) control. h, Fluorescent plate area of 1 nM oligomycin-treated cocultures of labeled clone N with unlabeled clone N or insensitive clone V (n = 3 samples) transfected with the indicated siRNA with or without the addition of exogenous asparagine. Error bars represent mean ± s.d.; * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 and **** P ≤ 0.0001 by two-tailed Mann–Whitney test (b), two-tailed Student’s t test (c and f) or one-way ANOVA with a Tukey post hoc test (d, e and h); alanine P < 0.0001, asparagine P = 0.0043, aspartate P = 0.0043, glutamate P = 0.0184, glycine P = 0.0013 and proline P < 0.0001 (b); P = 0.0008 (c); DMEM versus +asparagine P < 0.0001, +asparagine versus +glycine P = 0.0001), +asparagine versus +serine P = 0.0419, +asparagine versus +alanine P = 0.0008, +asparagine versus +proline P = 0.0001, +asparagine versus +aspartate P = 0.0016 and +asparagine versus +glutamate P = 0.0122 (d); DMEM versus aspartate 20 mM P = 0.0002 and aspartate 100 µM versus aspartate 20 mM P = 0.0004 (e); mock versus +asparagine for oligomycin P = 0.0011, phenformin P < 0.0001 and IACS-10759 P = 0.0006 (f); V siNT versus V siAsns P < 0.0001, V siAsns versus V siAsns + asparagine P = 0.0572 (h).

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