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. 2022 Dec 8;13:7548. doi: 10.1038/s41467-022-34622-w

Fig. 4. Parasite proliferation in blood and gonadal adipose tissue.

Fig. 4

a Cell-cycle analysis assayed by the number of Kinetoplasts (K) and Nuclei (N) in BSFs and ATFs Lister 427 parasites from mice infected for 5 days (n = 4 independent experiments). Left side: Representative image of in vivo microscopy of the adipose tissue and a vessel (scale bar, 20 µm); Right side: representative images of the three parasites analyzed on a monolayer (BSFs) and in vivo (ATFs) (Hoechst, blue; scale bar, 5 µm). b Percentage of BSFs (red circles, bars) and ATFs (yellow circles, bars) in each stage of the cell cycle. More than 800 cells per condition. Error bars represent the average and the standard deviation. c Ex vivo microscopy images of Lister 427 BSFs and ATFs labeled with EdU (green) and Hoechst (blue) (n = 5 independent experiments) (scale bar, 20 µm). Insets: representative images of labeled parasites (scale bar, 5 µm). d Percentage of EdU positive BSFs and ATFs. More than 300 cells per condition. Generalized linear mixed-effects model [GLMER], p = 0.00031. e Representative images of ex vivo microscopy of the inoculum parasites and a monolayer of BSFs stained with CTV and in vivo microscopy of ATFs and BSFs (scale bar, 20 µm). Insets: Representative images of labeled parasites (scale bar, 5 µm). f Distribution of the CTV Mean Fluorescence Intensity of BSFs and ATFs from mice infected for 2 days assessed by intravital microscopy. First bin of 20, remaining bins of 40 (n = 183 BSFs and n = 196 ATFs pooled from the 4 independent experiments). g Doubling time estimated by in vivo microscopy analysis of Lister 427 BSFs (red circles) and ATFs (yellow circles).(n = 183 BSFs and n = 196 ATFs pooled from the 4 independent experiments). Two-sided linear mixed-effects model [LME], p < 0.0001. Gray lines represent the average values and error bars the standard deviation. h Representative image of ex vivo microscopy of cultured Lister 427 parasites labeled with PKH26 (red) (scale bar, 10 µm) (n = 2 independent experiments). i, Representative flow cytometry profiles of non-labeled and PKH26 labeled cultured Lister 427 parasites and BSFs and ATFs isolated from mice infected for 3 and 4 days (n = 3 independent experiments) NC: Negative Control, D0: Day0, D3: Day3, D4: Day 4. Source Data are provided as a Source Data file.