Fig 2: Morphology observation of five novel Streptomyces phages. (A) Plaque morphologies of the five phages. Double agar overlays were performed to infect S. venezuelae ATCC 10712 with the phages Alderaan and Coruscant, and S. coelicolor M600 with the phages Dagobah, Endor1, and Endor2. Plates were incubated overnight at 30 °C and another day (3 days in the case of Dagobah) at room temperature to reach full maturity of the bacterial lawn. (B) Close-ups of phage plaques imaged using a stereomicroscope Nikon SMZ18. S. coelicolor M145 was infected by phages using GYM double agar overlays. The plates were incubated at 30 °C overnight and then kept at room temperature for two (Endor1 and Endor2) or three days (Dagobah). Scale bar: 1 mm. (C) Transmission electron microscopy (TEM) of phage isolates. The phage virions were stained with uranyl acetate. Scale bar: 150 nm.

Fig 3: Expression patterns of transgenic reporter lines.The 3xP3 promoter drives expression of transgenesis markers in eyes and nerve cells. a. Control 3xP3-RFP transgenic A. gambiae larva, image is a merge of green and red channels. Red arrows point to RFP expression in nerve cells, used as transgenesis marker (tm). b. TEP1-GFP, 3xP3-RFP larva. Arrows point to some of the lobes of the fat body expressing GFP. Note the fine architecture of thoracic fat body revealed by GFP expression. c. control wild type pupa, showing little auto fluorescence in the green channel. d. TEP1-GFP, 3xP3-RFP pupa showing GFP expression in fat body (fb, white arrows) and RFP expression in the eye (red arrows, tm). e-g. Larva carrying both ppo6-RFP, 3xP3-YFP and TEP1-GFP, 3xP3-RFP. RFP is expressed in hemocytes (white arrows) while GFP is expressed in fat body cells. YFP and RFP transgenic markers (red arrows, tm) are expressed in the nervous system. Scale bar = 200 µm h. RFP and YFP expression in ppo6-RFP, 3xP3-YFP pupa, showing RFP expression in hemocytes (hc). White arrows point to some RFP expressing hemocytes. Transgenesis YFP marker is expressed in the eye (red arrow, tm). i. Midgut of TEP1-GFP,3xP3-RFP female 24h after blood feeding, showing distinct GFP expression in proventriculus. (pv) j. Close-up on a proventriculus expressing GFP in TEP1-GFP gut. k. LRIM1-GFP,OpIE2-pac larva showing GFP expression in the fat body, alongside a control (wt) larva. l. Larva expressing APL1C-GFP, 3xP3-CFP showing GFP expression in the fat body (white arrows) and CFP transgenesis marker expression (red arrows, tm) in the nervous system (delineated with a thin line) m. APL1C-GFP,3xP3-CFP pupa expressing GFP in the fat body (white arrows) and CFP transgenesis marker in the eyes (tm, red arrow). n. TEP1-GFP (1) and APL1C-GFP (3) adult females with intense reporter expression in the fat body, visible through the cuticle, alongside a control female (2). Images a-h, j-m were acquired on a Zeiss Axiovert 200M microscope using a 5x or 10x objective. Images i and n were acquired on a Nikon SMZ18 binocular fluorescence microscope. Scale bar = 500µm except for d-h, l-m: 200 µm.

Fig 4: Lipoteichoic acid (LTA) from BPL1 strain demonstrates fat-reducing activity. A. Fat-reducing effect of purified LTA at different doses. B. Representative images of Nile red staining of lipid content in live young adult C. elegans in a wild-type N2 animal under fluorescence microscopy. Nematodes were fed with BPL1 cells, heat-treated BPL1 cells (HT-BPL1) or LTA (10 µg ml-1). Scale bar 250 µm. Original image taken by the authors for this paper with a Nikon-SMZ18 Fluorescence Stereomicroscope. C. Quantification of triglyceride content (mM TG/mg protein) in C. elegans fed with purified LTA from BPL1. D. Analysis of fat-reducing activity of LTAs from B. longum ES1 and B. animalis subsp. lactis BB12. For a, b percentage of fluorescence in bacterial and LTA-fed nematodes (wild-type strain N2) is represented. Nile red was quantified at young adult stage. Orlistat (6 µg ml-1) was used as positive control. Data are mean ± SD. and were calculated from two biological independent experiments. For A, B, C, D *P < 0.05; **P < 0.01; ***P < 0.001; NS, not significant.

Fig 5: Induced fluorescent and dark pseudopupil overlap. (a) Head of the bee Anthophora sp. These bees have clear eyes and its dark pseudopupil is evident as a black spot on the eye surface. Image acquired with a Nikon SMZ18 equipped with a bright field filter cube (P2-EFLBF) and a ¼ wave plate. The eye was stained with Neurobiotin 488 (see “Methods”). Scale bar: 500 µm. (b) Zoomed-in view of the area marked by the white rectangle in (a) showing more clearly the dark pseudopupil in the left eye. It is common to see multiple pseudopupils resulting from the incident light from the environment around the eye. (c) Zoomed-in view of the area marked by the white rectangle in (a), after using a P2-EFLGFP-B filter cube—the induced fluorescent pseudopupil was visible in the same sample (in vivo). (d) Co-visualization of the dark (b) and induced fluorescent pseudopupil (c) showing a clear overlap of the two. (b–d) Scale bar: 200 µm. (e) Relative pixel values for the induced fluorescent and dark pseudopupils in the area marked in (d) by a white rectangle. Note that the centres of the two (i.e. max relative pixel value) coincide, with the induced fluorescent pseudopupil having the centre much more easily identifiable compared to the dark pseudopupil.