Fig. 4: Constitutively active YAP (YAP 5SA) expression rescues YAP transcriptional activity and glycolysis in Q cells. | Nature Communications

Fig. 4: Constitutively active YAP (YAP 5SA) expression rescues YAP transcriptional activity and glycolysis in Q cells.

From: Metabolic and transcriptomic reprogramming during contact inhibition-induced quiescence is mediated by YAP-dependent and YAP-independent mechanisms

Fig. 4

a Luciferase assays with the 8×GTIIC-Lux reporter in WT and 5SA cells in the P and Q states. Data are normalized to WT P cells and are presented as the mean ± SEM of 4–5 biologically independent samples. P values by unpaired two-tailed student’s t-test are indicated except for ****P < 0.0001. b, c The extracellular acidification rate (ECAR) and individual glycolysis parameters in WT and 5SA cells in the P and Q states. Data are presented as the mean ± SEM of 3–4 biologically independent samples. P values by unpaired two-tailed student’s t-test are indicated. Glc glucose, Oligo oligomycin, 2DG 2-deoxyglucose. d Immunoblots and protein quantification of glycolytic enzymes and p27kip1 in WT and 5SA cells in the P and Q states. Vinculin was monitored as a loading control. Representative immunoblots of four independent experiments are shown. Values are the mean ± SEM of 3-4 biologically independent experiments. P values by unpaired two-tailed student’s t-test are indicated except for ****P < 0.0001 and ns (PFK1 P = 0.0667, ALDOA P = 0.1451, P p27kip1 P = 0.7940, Q p27kip1 P = 0.9048). HK hexokinase, PFK1 phosphofructokinase 1, PKM pyruvate kinase M, ALDOA aldolase A, LDHA lactate dehydrogenase A. e Comparison of the quantified key extracellular metabolic fluxes (estimated flux ± SD) in WT and 5SA cells in the P and Q states. The flux rates of glucose uptake and lactate excretion are plotted. P values by unpaired two-tailed student’s t-test are indicated in the figure except for ****P < 0.0001 and ns P values (Glucose Cons. Q WT P = 0.3440, Lactate Pro P 5SA P = 0.0557) f Metabolic flux maps of WT and 5SA cells in the P and Q states. The flux maps were determined using 13C-MFA of multiple datasets at isotopic steady state as described in Methods. g Comparison of key intracellular glycolytic fluxes (estimated flux ± SD) in WT and 5SA cells in the P and Q states. All flux fits passed the chi-square goodness-of-fit test. P values by unpaired two-tailed student’s t-test are indicated except for ns P = 0.5742.

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