Fig. 2: Vaha is synthesized in the gut and not in the brain. | Nature Communications

Fig. 2: Vaha is synthesized in the gut and not in the brain.

From: A nutrient responsive lipase mediates gut-brain communication to regulate insulin secretion in Drosophila

Fig. 2

a RT-qPCR analysis of Vaha mRNA level in gut and CNS samples isolated from w1118. Vaha expression is normalized to GAPDH. n = 3, 10 guts and 10 CNS per replicate. Data is assessed by two-tailed Student’s t-test and presented as mean ± SD. b Vaha RNAi in the gut (w-; NP1 Gal4/Vaha V5; Vaha RNAi/gd2HF), neurons (w-; Elav Gal4/Vaha V5; Vaha RNAi/gd2HF) fat body (w-; Ppl Gal4/Vaha V5; Vaha RNAi/gd2HF) and imaging of Vaha V5 in the IPCs. Control flies are w-; NP1 Gal4/Vaha V5; +/gd2HF, w-; Elav Gal4/Vaha V5; +/gd2HF and w-; Ppl Gal4/Vaha V5; +/gd2HF. Confocal images are projection of z stacks, n = 6 brains, scale bar 25 μm. c Quantification of Vaha V5 intensity in the IPCs of Vaha RNAi in the gut. Each data point represents fluorescence intensity in one IPC, n = 49 IPCs (control), n = 46 IPCs (RNAi). Data is assessed by two-tailed Student’s t-test and horizontal line indicates median. d Quantification of Vaha V5 intensity in the IPCs of neuronal knockdown of Vaha. Each data point represents fluorescence intensity in one IPC, n = 43 IPCs (control), n = 45 IPCs (RNAi). Data is assessed by two-tailed Student’s t-test and horizontal line indicates median. e Quantification of Vaha V5 intensity in the IPCs of fat body knockdown of Vaha. Each data point represents fluorescence intensity in one IPC, n = 46 IPCs (control), n = 40 IPCs (RNAi). Data is assessed by two-tailed Student’s t-test and horizontal line indicates median. f Live imaging of gut and corresponding brain dissected from flies expressing UAS nuclear localized mCherry driven by Vaha Gal4 driver (w-; Vaha Gal4/+; UASmcherry.NLS/+). n = 6 guts and brains. Scale bar gut 450 μm, brain 300 μm. g Immunostaining of guts dissected from above flies. Colored brightfield image highlights the midgut in blue and hindgut in brown. R1-R5 subdivisions of the midgut are marked, MT are malpighian tubules. mCherry is pseudocolored orange while Cut antibody marking R3 is pseudocolored red. n = 6 guts, Scale bar 200 μm. Source data are provided for 2a, c, d, e as a source data file.

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