Fig 2: Colony growth following highthroughput robotic pinning demonstrates budding and hyphal morphologies of Z. tritici. Displayed are images of technically duplicated spots of the Z. tritici ?ku70 strain HLS1000 following 6, 12 and 16 days growth at 18° on ZTSC medium. Approximately 1000 cells in a volume of 2 µl were pinned onto agar. Colonies consisted of a budding mucoid morphology surrounded by hyphae at the periphery. Images were captured using a Leica M205FA stereo microscope. Scale bars are 1 mm length.

Fig 3: NFPs regulate transmural growth and cardiomyocyte proliferation autonomously. (A) Control hearts and (B) iDKO hearts show sporadic clones labeled with different colors. Cardiomyocytes in hearts were induced to be labeled at E7.75 with dosage of tamoxifen at 20 µg/g body weight and the hearts at E12.5 were imaged via Leica M205 FA, which can detect fluorescent protein-labeled cells at single-cell resolution. (C–E) Individual clones in the cleared hearts were 3D-imaged to identify the geometric pattern of each of the clones. C, D, and E show compact, trabecular and transmural clones, respectively. (F) The clonal patterns between control and iDKO. (G) The transmural growth of the clones and that the iDKO clones grow significantly shorter distances compared with the control clones. (H) The number of cells in each clone; the iDKO clones contain significantly more cells than the control clones across all 3 types of clones. (Scale bars in A and B: 200 µm and in C–E: 50 µm.)

Fig 4: Macrophages respond more prominently than neutrophils to T. carassii infection. Tg(mpeg1:mCherry-F;mpx:GFP) were injected intravenously at 5 dpf with n = 200 T. carassii or with PVP. At 4 dpi, larvae were separated in high- and low-infected individuals. (A–B) At the indicated time points, images were acquired with Leica M205FA Fluorescence Stereo Microscope with 1.79x zoom. Images are representatives of n = 5–47 larvae per group, depending on the number of high- or low-infected larvae categorised at each time point, derived from two independent experiments. Scale bar indicates 750 µm. (C) Corrected Total Cell Fluorescence (CTCF) quantification of infected and non-infected larvae. Owing to the high auto-fluorescence, the gut area was excluded from the total fluorescence signal as described in the Materials and methods section. Bars represent average and standard deviation of red and green fluorescence in n = 5–47 whole larvae, from two independent experiments. * indicates significant differences (p<0.05) to the respective PVP control as assessed by Two-Way ANOVA followed by Bonferroni post-hoc test. Figure 3—source data 1.Macrophages respond more prominently than neutrophils to T. carassii infection.

Fig 5: SMAR mESCs contribute to form chimeras (F0), but the genetic modification does not affect the progeny (F1)(A) Chimeras generated with pSMAR (clone v71c22) or nSMAR (clone v85c17) mESCs. Between 6 and 12, stably transfected mESCs were microinjected into C57BL/6NxB6D2F1 embryos, resulting in the formation of chimeras, as observed by the agouti/chinchilla coat color.(B) Transgenic GFP expression of ear biopsies (n = 49) at the time of weaning compared with C57BL/6N control mice (n = 3). The MFI from fluorescent images is expressed as relative light units (RLU). The statistical analysis was performed using an unpaired T test with Welch's correction (****p < 00.001).(C) Comparison of fluorescence from ear biopsies of pSMAR (n = 23) and nSMAR (n = 26) chimeras with control mice (n = 3). The statistical analysis was performed using a Brown-Forsythe and Welsch ANOVA with Bunnett's T3 multiple comparison test (***p = 00.004; ****p < 00.001).(D) Summary of transgene expression from representative chimeric organs. See Table S3 for a complete dataset.(E) SC contribution to the germline of two male chimeric mice generated with mESCs engineered with nSMAR (mouse 4) and pSMAR (mouse 5). The males were backcrossed with C57BL/6J females and generated 100% agouti litters.(F) Genotyping PCR from tail biopsies of chimeric mice (F0) and their respective litters (F1). C57BL/6 was used as a negative control. The amplicon corresponds to a 515 bp GFP band. An internal mammalian-conserved SOX21 sequence (237 bp) was used as an internal control.(G) PCR amplification of the transgene before (germ cells) and after (sperm) meiosis. The GFP amplicon corresponds to the 515 bp band. An internal mammalian conserved SOX21 sequence (237 bp) was used as an internal control.(H) Fluorescent images and FACS analysis depicting transgene expression across gametogenesis. GFP fluorescent testis of mouse 4 (nSMAR) and mouse 5 (pSMAR). Constitutively expressing UBC:GFP mouse (Jackson lab, Pos Ctrl) and a C57BL/6N mouse (Neg Ctrl). (Leica M205FA, exposure 1s, amp gain 1.9x, digital exposure 4, scale bars = 2 mm and 50 µm). FACS analysis was performed in germinal cells from digested seminiferous tubes. The sperm was collected and imaged using a Nikon Ti microscope. Cartoon adapted from (Falcone and Hurd, 2007).(I) Immunohistochemistry of seminiferous tubules' sections (left = GFP staining with hematoxylin counterstaining, middle = unstained control, right = hematoxylin-eosin staining, scale bars = 100 µm).