Additional file 1: Figure S1. of Further characterization of adult sheep ovarian stem cells and their involvement in neo-oogenesis and follicle assembly

The presence of two stem cell population in OSE by flow cytometry analysis is shown. Table S1. List of antibodies used in the study. Table S2. List of primers used in the study. Figure S2. H & E staining of freshly isolated sheep OSE smears (arrows represent small putative VSELs smaller then RBCs and asterisk represent OSCs and cell doublets (circled) show dividing stem cells). Figure S3. RT-PCR on freshly isolated OSE cells and granulosa cells surrounding oocytes. Figure S4. FSHR expression on ovarian stem cells. Figure S5a. Z stack of OCT-4 expressing germ cell clusters in FSH treated OSE cell culture. Figure S5b. Z stack of FSHR expressing germ cell clusters in FSH treated OSE cell culture. Figure S6. Co-expression of FSHR and SSEA-4 on ovarian stem cells. Figure S7. FSHR expression on proliferating and dividing ovarian stem cells cultured in vitro within OSE cells on FSH treatment. Figure S8. H&E staining of ovarian sections (A) shows a distinct layer of OSE cells (B-C) PCNA immuno-localization on sheep ovarian sections showed few cells in OSE positive for PCNA, cluster of oocytes/germ cell cyst, individual primordial and primary follicles located in cortical region of ovary showed strong nuclear PCNA with faint stain in ooplasm. Figure S9a. Negative control for ICC to show specificity of staining (i) Negative for VASA (ii) Negative for SSEA-4 and PCNA (iii) Negative for FSHR and OCT-4. Figure S9b. Negative control by peptide blocking of FSHR antibody to show specificity of staining obtained using FSHR. (PDF 1911 kb

Additional file 1: of Delineating the effects of 5-fluorouracil and follicle-stimulating hormone on mouse bone marrow stem/progenitor cells

Description of methods, primers and antibodies used in the study. (PDF 340 kb

Additional file 6: of Transcriptional activator DOT1L putatively regulates human embryonic stem cell differentiation into the cardiac lineage

ChIP sequencing of occupancy of H3K79me2 on DMD gene during cardiac differentiation of KIND1 and HES3 cells showing occupancy of H3K79me2 methylation mark brought about by DOT1L on DMD gene during cardiac differentiation. Results clearly show significant peaks representing the DOT1L specific methylation mark on days 12 and 20 as compared to day 0 suggestive of its activation by DOT1L during cardiac differentiation in vitro. (PDF 614 kb

Additional file 3: of Transcriptional activator DOT1L putatively regulates human embryonic stem cell differentiation into the cardiac lineage

Characterization of cardiac differentiation of HES3 cells by immunofluorescence studies. Expression of NKX2.5 (A) and CTNT (B) on days 12 and 20 observed by immunofluorescence. (A) Distinct nuclear expression of NKX2.5 observed and (B) CTNT cell surface expression. Similar changes observed when KIND1 cells were differentiated into cardiac cells as described earlier [43]. Counterstaining using DAPI. Magnifications 20×. (PDF 450 kb

Additional file 2: of Transcriptional activator DOT1L putatively regulates human embryonic stem cell differentiation into the cardiac lineage

Characterization of cardiac differentiation of HES3 cells by quantitative real-time PCR (qRT-PCR). Expression of transcripts representing pluripotency (OCT4), cardiac mesoderm (MESP1), cardiac progenitors (NKX2.5, MEF2C), and cardiomyocytes (CTNT) at days 0, 12, and 20 during 20 days of cardiac differentiation. Note OCT-4 expression in undifferentiated cells is downregulated as the cells initiate differentiation. Early cardiac markers detected on day 12 and mature markers upregulated on day 20. Similar changes in transcripts expression observed when KIND1 cells were differentiated into cardiac cells as described earlier [43]. Error bars represent ÂąSEM. (PDF 410 kb

Additional file 5: of Transcriptional activator DOT1L putatively regulates human embryonic stem cell differentiation into the cardiac lineage

Dystrophin gene expression during cardiac differentiation of KIND1 hES cells on days 0, 12, and 20 during cardiac differentiation of KIND1 hES cell line. Expression of Dystrophin increased in cardiac progenitors and cardiomyocytes compared to undifferentiated KIND1 cells. Results in agreement with earlier reports in DOT1L conditional knockout mice heart concluding Dystrophin as a direct target of DOT1L [35]. Error bars represent ¹SEM. (PDF 329 kb

Additional file 1: of Transcriptional activator DOT1L putatively regulates human embryonic stem cell differentiation into the cardiac lineage

Brightfield images of HES3 hES cells during directed differentiation into cardiac lineage. Differentiation results in distinct morphological changes leading to increased compaction among the cells as differentiation proceeds from day 0 to day 20. Similar changes observed when KIND1 cells were differentiated into cardiac cells as described earlier [43]. Magnification 10×. (PDF 554 kb

Additional file 4: of Transcriptional activator DOT1L putatively regulates human embryonic stem cell differentiation into the cardiac lineage

ChIP sequencing in KIND1 and HES3 cells during cardiac differentiation visualized by Integrated Genome Viewer shows binding profile of H3K79me2 modification across genes HNF4A, LEFTY1, NOGGIN, NQO1, OTX2, and NPTX2 in KIND1 (green) and HES3 (red) cells at days 0, 12, and 20 of cardiac differentiation. (PDF 548 kb