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. 2020 Jun 12;9:e53247. doi: 10.7554/eLife.53247

Figure 5. FAO loss in hemocyte progenitors led to sustained glycolysis.

(A) ATP levels in control and whd1/whd1 whole larvae. p-Value of whd1/whd1compared to control = 5.327×10−2. (B–D) Glucose incorporation (marked by 2-NBDG uptake) levels in control dome-MESO-EBFP2/+ (B–B'') and dome-MESO-EBFP2/+; whd1/whd1 (C–C'') lymph glands. (D). Quantitative analysis of results from B–C demonstrating a significant increase in glucose uptake in the whd1/whd1 progenitors. p-Value for dome-MESO-EBFP2/+; whd1/whd1 = 6.09×10-7 compared to control. (E–F) Increased lactate dehydrogenase in-vivo enzymatic staining assay of whd1/whd1 lymph gland (F) compared to control (E). (G) Fold change in the level of Hex-A mRNA expression in control w1118 and whd1/whd1 lymph glands. p-Value of whd1/whd1 = 6.379×10−3 compared to control. (H) Fold change in the level of Pfk mRNA expression in control w1118 and whd1/whd1 lymph glands. p-Value of whd1/whd1 = 3.739×10−3 compared to control. (I–M) Proliferation status (marked by EdU) in control dome > GFP (I–I'), dome > GFP; whd1/whd1 (J–J'), 2-DG fed dome > GFP; whd1/whd1 (K–K') and dome > GFP; whd1/whd1; UAS-Glut1 RNAi (L–L') lymph glands. (M). Quantitative analysis of results from I–L'. p-Value for dome > GFP; whd1/whd1 = 4.37×10−7 compared to control and p-value for dome > GFP; whd1/whd1 = 3.25×10−7 fed with 2-DG compared to non-fed dome > GFP; whd1/whd1. p-Value for dome > GFP; whd1/whd1; UAS-Glut1 RNAi = 4.53×10−7 compared to non-fed dome > GFP; whd1/whd1. (M–P) Comparison of differentiation (marked by P1) levels in control dome > GFP (M), dome > GFP; whd1/whd1 (N) and 2-DG fed dome > GFP; whd1/whd1 (O) lymph glands. (P). Quantitative analysis of results from M–O show decline in proliferation upon 2-DG feeding. p-Value for dome > GFP; whd1/whd1 = 4.43×10−11 compared to control and p-Value for dome > GFP; whd1/whd1 = 8.6×10−2 fed with 2-DG compared to non-fed dome > GFP; whd1/whd1. p-Value for dome > GFP; whd1/whd1; UAS-Glut1 RNAi = 5.9×10−2 compared to non-fed dome > GFP; whd1/whd1. n.s. = not significant. Individual dots represent biological replicates. Values are mean ± SD, asterisks mark statistically significant differences (*p<0.05; **p<0.01; ***p<0.001, Student’s t-test). Scale bar: 20 µm.

Figure 5—source data 1. Contains numerical data plotted in Figure 5A,D,G,H,M and R.

Figure 5.

Figure 5—figure supplement 1. 2-NBDG assay in the lymph gland primary lobe.

Figure 5—figure supplement 1.

(A–B) Glucose incorporation (marked by 2-NBDG uptake) levels in control dome-MESO-EBFP2/+; Hml-DsRed lymph glands (A–A'''). (B) Quantitative analysis of results from A–A'''. p-Value for IPs = 3.64×10−4 compared to progenitors. p-Value for Cortical Zone Hml+=1.83×10−6 compared to progenitors. Individual dots represent biological replicates. Values are mean ± SD, asterisks mark statistically significant differences (*p<0.05; **p<0.01; ***p<0.001, Student’s t-test). Scale bar: 20 µm.
Figure 5—figure supplement 1—source data 1. Contains numerical data plotted in Figure 5—figure supplement 1B.