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Fig. 4 | BMC Biology

Fig. 4

From: Peripheral cathepsin L inhibition induces fat loss in C. elegans and mice through promoting central serotonin synthesis

Fig. 4

Knockdown of cpl-1 in the intestine and hypodermis promoted serotonin synthesis in ADF neurons. a Representative images and quantification of Oil Red O staining in tissue-selective RNAi strains: intestine-restricted VP303 (rde-1(ne219) V;kzls7), hypodermis-restricted NR222 (rde-1(ne219) V;kzls9), germline-restricted MAH23 (rrf-1(pk1417) I), muscle-restricted WM118 (rde-1(ne300) V;nels9 X), and neurons-restricted VH624 (rhIs13 V; nre-1(hd20) lin-15B(hd126) X), during control RNAi or cpl-1 RNAi treatment. The data were obtained from 3 independent experiments, and 3000 stained worms were used in each experiment. b, c Real-time PCR analysis of genes involved in the serotonin signaling pathway in intestine-selective cpl-1 knockdown worms (b) and hypodermis-selective cpl-1 knockdown worms (c). act-1 was used as the reference gene in real-time PCR analysis, n = 3 independent growths. d The serotonin levels in intestine-selective (VP303) and hypodermis-selective (NR222) cpl-1 knockdown worms; the data were based on 3 independent experiments. e The expression of tph-1::gfp in systemic, intestine-selective (VP303), and hypodermis-selective (NR222) cpl-1 knockdown worms; tph-1 expression was visible in NSM (arrowheads) and in ADF (arrows). For GFP fluorescence quantification, the data were obtained from 3 independent experiments and 30 worms were imaged and qualified with the level of fluorescence intensity. f The pharyngeal pumping rate in intestine-selective (VP303) and hypodermis-selective (NR222) cpl-1 knockdown worms; the data were obtained from 3 independent experiments, and 30 worms were used in each experiment. g The two-choice olfactory preference assays to PA14 and HT115 bacteria in intestine-selective (VP303) and hypodermis-selective (NR222) cpl-1 knockdown worms, n = 3 independent growths. All data in ad, f, and g are presented as mean ± SEM. Two-tailed Student’s t test was used for statistical analysis. *p < 0.05; **p < 0.01; ***p < 0.001. n.s., not significant

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