Fig. 1.

Glycolytic enzymes and glucose flux in Mec-1 cells. 10 [4] Mec-1 cells were treated with 10 μM P-M2tide or 10 μM DASA, respectively (both with no cytotoxicity towards Mec-1 cells), and with fludarabine (simultaneously, under hypoxia for 24 h, n = 4; A). Viability was defined as % luminescence of the untreated control. Lactate efflux after 24 h was measured using 10⁶ Mec-1 cells (n = 2; B). 5-³H-glucose turn-over was assessed after treatment with PM2-tide or DASA (under hypoxia, 24 h, n = 3; C). For qRT-PCR fold change of expression (compared to untreated control) of glycolytic Iso−/enzymes Hexokinase (HK), Glucose-6-phhosphate isomerase (GPI), Phosphofructokinase (PFKL, two different primers), Aldolase A/B/C (ALDOA, ALDOB and ALDOC each two different primers), Triose-phosphate isomerase (TPI), Phosphoglycerate mutase (PGM), Enolase (ENO), Pyruvate kinase (PKLR two different primers (PKLR was not detectable with primer #1); PKM2) and Lactate dehydrogenase (LDHA, LDHC with two primers (#1 and #2) and repetitive testing) was measured in untreated cells and after treatment with P-M2tide or DASA (under hypoxia, 24 h, n = 3; D). A minimum of 2-fold change (relative quantification of more than two or <0.5; dashed lines) was considered significant. Error bars are indicating standard deviation. Statistical significance was calculated with a two-way t-test. Significance is represented as * for p-values <.05.