Fig 2: Zn tolerance phenotypes of wild-type yeast expressing AhPDF1.1b (A) or zrc1 mutant expressing AhPDF1.1b (B). (A.1) Drop-test assays of wild-type yeast expressing the pYX212 empty vector (WT-EV) or wild-type yeast expressing AhPDF1.1b (WT-AhPDF1.1b) on 20 mmol/L ZnSO4 agar plates. An overnight yeast culture was diluted to the A600nm levels indicated and 10 µL were spotted on YNB medium supplemented or not with 20 mmol/L ZnSO4. (A.2) Growth curves of wild-type yeast expressing the pYX212 empty vector (WT-EV) or wild-type yeast expressing AhPDF1.1b (WT-AhPDF1.1b) plated on agar medium supplemented with 20 mmol/L ZnSO4. Isolated colonies plated on 20 mmol/L ZnSO4 were photographed with a SZX16 Olympus stereozoom microscope and their areas were measured using ImageJ software. Error bars correspond to standard errors, n = 50. (A.3) Number of isolated colonies (CFU) of wild-type yeast expressing the pYX212 empty vector (WT-EV) or wild-type yeast expressing AhPDF1.1b (WT-AhPDF1.1b) plated on agar medium supplemented with 20 mmol/L ZnSO4. The number of colonies was counted after plating 100 µL of a A600nm = 0.01 culture of yeast cells. The Student's t-tests give a P = 0.47 on the control medium and a P = 0.27 on the zinc-supplemented medium. Drop-test assays of zrc1 yeast expressing the empty vector (zrc1-EV) or zrc1 yeast expressing AhPDF1.1b (zrc1-AhPDF1.1b) on 10 mmol/L ZnSO4 agar plates. An overnight yeast culture was diluted to the A600nm levels indicated and 10 µL were spotted on YNB medium supplemented or not with 10 mmol/L ZnSO4. B.2 Growth curves of zrc1 yeast expressing the empty vector (zrc1-EV) or zrc1 yeast expressing AhPDF1.1b (zrc1-AhPDF1.1b) obtained by measuring the areas of isolated colonies plated on 10 mmol/L ZnSO4. Error bars correspond to standard errors, n = 50. B.3 “Zinc shock” assay in liquid culture of zrc1 yeast expressing the empty vector (zrc1-EV) or zrc1 yeast expressing AhPDF1.1b (zrc1-AhPDF1.1b). The growth curves are followed by A600nm measurements during a zinc shock assay as described in [27]. Error bars correspond to standard errors, n = 3.

Fig 3: Parbendazole shows good anti-tumor activity in vivo. (A) is the tumor volume of mice; (B) is the body weight of mice. The xenograft tumor mice were randomly divided into four groups (n = 5 for each group) and given different doses of CA-4, parbendazole or solvent, once a day by intragastric administration. The tumor volumes were measured by an electronic caliper, and the body weight of mice were measured by an electronic balance. Results are expressed as mean ± SD; (C) indicates the therapeutic effect of papendazole on zebrafish bearing xenografts. The zebrafish bearing xenografts were randomly divided into four groups (n = 30 for each group) and were administered different doses of CA-4, parbendazole, or solvent alone. Images were obtained by an OLYMPUS SZX16 microscope, and the fluorescent integrated density was measured by Image J software. The scale bar is 100 µmol/L; (D) is the fluorescent integrated density of each group of zebrafish. Graphs were draw by GraphPad Prism 7.00. Results are expressed as mean ± SD (****P < 0.0001).

Fig 4: The efficiency of ndgr1a MO1 and MO2 was verified in vivo.(A–D) Fertilized Xenopus eggs were co-injected with 3.5 picomoles of coMO or ndrg1a MO1 in combination with an mRNA containing MO1 binding site followed by GFP coding sequence (MO1BS-GFP) and evaluated for live GFP translation at stage 18 with an Olympus SZX16 fluorescence microscope. (A, B) GFP. (C, D) Bright field view. (E–H) ndrg1a MO2 specifically inhibited the translation of GFP following its binding site (MO2BS-GFP), as revealed by the in vivo assay as above. (E, F) GFP. (G, H) Bright field view.

Fig 5: Effects of dsISWI and dsBRM on Tuta absoluta ovary development. (A) Ovaries of uninjected 4 d females. (B) Abnormal ovaries of 4 d females injected with dsISWI. (C) Abnormal ovaries of 4 d females injected with dsBRM. The red arrow represents the mature oocytes. The ovaries were photographed under an Olympus stereomicroscope (SZX16; Olympus, Tokyo, Japan). See more pictures in Figure S5.