Fig 1.
Effect of JQ-1 on male-female pairing rate, egg production, and egg morphology in S. japonicum.
Effects of different concentrations (5 μM, 10 μM, and 15 μM) of JQ-1 application on male-female pairing stability (A), egg production (B), and egg morphology (C-F) in S. japonicum pairs cultured in vitro for 10 d. Data represent the mean ± SEM of three independent experiments. Scale bars: 200 μm. Asterisks show statistical differences (***P < 0.001) tested by one-way ANOVA with multiple comparisons (Tukey’s post-hoc test).
Fig 2.
Effect of JQ-1 on cell proliferation in male-female pairs of S. japonicum.
Red signals indicate active mitotic cells labeled by EdU; blue signals, Hoechst-positive cells. (A-D) Male S. japonicum and (E-H) female S. japonicum. EdU-incorporated cells are detected in the testes and parenchyma of untreated males (A) and in the vitellarium and ovary of untreated females (E). EdU-positive cells are detected after application of JQ-1 at 5 μM (B, F), 10 μM (C, G), and 15 μM (D, H). Abbreviations: ov, ovary; t, testes; SV, seminal vesicles. Scale bars: 200 μm.
Fig 3.
Morphological changes of spermatozoa in testes and seminal vesicles of S. japonicum treated with JQ-1 in vitro.
Worms were stained with carmine hydrochloride and analyzed using confocal laser scanning microscopy. (A, E) Control worms; worms treated with JQ-1 at 5 μM (B, F), 10 μM (C, G), and 15 μM (D, H). (A-D) Scale bars: 20 μm; (E-H) Scale bars: 10 μm. Abbreviations: t, testis; SV, sperm vesicle. (I) Comparison of the diameter of the testicular lobes after JQ-1 application at the indicated concentration for 10 d. Data represent the mean ± SEM (n ≥ 15 for each group). Asterisks show statistical differences (**P < 0.01; ***P < 0.001) tested by one-way ANOVA with multiple comparisons.
Fig 4.
Morphological changes of ovaries and yolk glands in female S. japonicum treated with JQ-1.
Worms were stained with carmine hydrochloride and analyzed using confocal laser scanning microscopy. (A, E) Control worms; worms treated with JQ-1 at 5 μM (B, F), 10 μM (C, G), and 15 μM (D, H). Abbreviations: ov, ovary; v, vitellarium; (A-H) Scale bars: 20 μm. Comparison of the length, width, and area of the ovary after JQ-1 application at the indicated concentration (I-K) for 10 d. Data represent the mean ± SEM (n ≥ 15 for each group). Asterisks show statistical differences (**P < 0.05, **P < 0.01, ***P < 0.001) tested by one-way ANOVA with multiple comparisons.
Fig 5.
Results of quantitative PCR analyses of S. japonicum cultured with or without JQ-1 for 10 d.
Relative transcription level of Nanos1 in male (A) and female S. japonicum (B). Relative transcription level of Plk1 in male (C) and female (D) S. japonicum. Relative transcription level of Brd2 in male (E)€ and female (F) S. japonicum. Data represent the mean ± SEM of three independent experiments. Asterisks show statistical differences (*P < 0.05; **P < 0.01) tested by one-way ANOVA with multiple comparisons.
Fig 6.
Effect of JQ-1 treatment on liver granuloma in mice infected with S. japonicum.
(A) Protocol used to assess liver granuloma in mice. (B) Gross appearance of livers obtained from mice infected with S. japonicum and treated with JQ-1 or vehicle (HP-β-CD). Liver slices stained with hematoxylin and eosin. Scale bars: 500 μm. (C) Measurement of granuloma area as a percentage of total area as assessed by computer-aided morphometry. (D) Liver weights of S. japonicum infected mice treated with JQ-1 or HP-β-CD. (E) Spleen weights of S. japonicum infected mice treated with JQ-1 or HP-β-CD. (F) Effect of JQ-1 treatment on serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in mice infected with S. japonicum. (G) Effect of JQ-1 treatment on mRNA expression of inflammation-related genes in the liver of mice infected with S. japonicum. Data represent the mean ± SEM (n = 9 for each group). Asterisks denote statistically significant differences (Student’s t-test, *P < 0.05; **P < 0.01) vs. the HP-β-CD–treated control group.
Fig 7.
JQ-1 treatment alters germ cell proliferation of S. japonicum and egg production in the liver of mice infected with S. japonicum.
(A) Egg morphology and (B) production in the liver. (C) Numbers of adult worms and (D) worm pairs in the liver. (E-F) Results of quantitative PCR analyses of S. japonicum detected in S. japonicum–infected mice treated with JQ-1. (G) Red signals indicate active mitotic cells labeled by EdU; blue, Hoechst-positive cells. EdU-incorporated cells in control worms were detected in the testes and parenchyma of males and in the vitellarium and ovary of females. (A) Scale bars: 500 μm. (G) Abbreviations: ov, ovary; t, testes. Scale bars: 100 μm. Data represent the mean ± SEM (n = 9 for each group). Asterisks denote statistically significant differences (Student’s t test, ** P < 0.01).
Fig 8.
Morphological changes in the testis and ovary of S. japonicum treated with JQ-1 in vivo.
Worms were stained with carmine hydrochloride and analyzed using confocal laser scanning microscopy. (A-C) Testes, seminal vesicles, and ovary of worms in control mice. (D-F) Testes, seminal vesicles and ovary of worms in mice treated with JQ-1. Abbreviations: ov, ovary; t, testes; SV, seminal vesicles. Scale bars: 20 μm.