Journal:

Article Title: CUL-4A stimulates ubiquitylation and degradation of the HOXA9 homeodomain protein

doi: 10.1093/emboj/cdg577

Figure Lengend Snippet: Fig. 6. CUL-4A ablates the HOXA9-induced differentiation block of 32Dcl3 myeloid progenitor cells. (A) Western blotting to detect stable expression of HOXA9 and MYC-CUL-4A in 100 µg of 32Dcl3, 32D-HOXA9, 32D-HOXA9/pcDNA3 and 32D-HOXA9/MYC-CUL-4A (clones 6-16 and 7-1) extracts using antibodies against HOXA9, MYC tag or β-actin. (B) Wright–Giemsa staining of the indicated 32Dcl3 cell lines grown in the presence of 5 ng/ml of IL-3, or induced to differentiate with 20 ng/ml of G-CSF for 8 days. Hemocytograms of the resulting 32Dcl3 cell lines are indicated on the right. The percentages of myoblast, intermediate and granulocytic cells were obtained by counting a total of 500 cells in each experiment, and data are presented as mean ± SD from four independent experiments. (C) The indicated 32Dcl3 cell lines were either untreated or treated with 20 ng/ml of G-CSF for 8 days, and analyzed by flow cytometry for expression of MAC1 (mCD11b) as a marker for granulocytic differentiation. Histograms showed the quantitation of MAC1–FITC+ staining and M1 values in each column indicate the percentages of differentiated (MAC1-positive) 32Dcl3 cells.

Article Snippet: The mononuclear cells at the interphase were collected, and were used to purify CD34 + cells using the immunomagnetic MACS CD34 Isolation kit (Miltenyi Biotec Inc.) according to the manufacturer’s instructions.

Techniques: Blocking Assay, Western Blot, Expressing, Clone Assay, Staining, Flow Cytometry, Marker, Quantitation Assay

Journal:

Article Title: CUL-4A stimulates ubiquitylation and degradation of the HOXA9 homeodomain protein

doi: 10.1093/emboj/cdg577

Figure Lengend Snippet: Fig. 2. CUL-4A is expressed in primitive and differentiated hematopoietic cells. (A) An indirect immunofluorescence assay was carried out on the indicated cells using the anti-CUL-4A polyclonal antibody and secondary antibody conjugated to Cy3 (top). The same cells were also stained with DAPI and photographed under phase contrast lenses (bottom). (B) Subcellular localization of CUL-4A in primary CD34+ hematopoietic stem/progenitor cells by immunofluorescent confocal microscopy. CD34+ cells were also stained with the TO-PRO-3 dye for visualization of nuclei.

Article Snippet: The mononuclear cells at the interphase were collected, and were used to purify CD34 + cells using the immunomagnetic MACS CD34 Isolation kit (Miltenyi Biotec Inc.) according to the manufacturer’s instructions.

Techniques: Immunofluorescence, Staining, Confocal Microscopy

Journal: The Journal of Clinical Investigation

Article Title: Liver sinusoidal endothelial cell progenitor cells promote liver regeneration in rats

doi: 10.1172/JCI58789

Figure Lengend Snippet: LSECs were isolated by elutriation, and the CD133+ fraction was obtained by immunomagnetic separation to obtain resident SPCs. (A) Staining for CD45 (FITC; green). (B) Staining for CD31 (Alexa Fluor 405; blue). (C) The merged image of A and B demonstrates that these CD133+ cells are also CD45+CD31+. Original magnification, ×100. (D) Scanning EM demonstrates the resident SPCs have fenestration organized in sieve plates characteristic of LSECs. Scale bar: 5 μm.

Article Snippet: Microbeads from the following kits were used: CD133 Cell Isolation Kit and Anti-FITC MultiSort Kit (Miltenyi Biotec).

Techniques: Isolation, Immunomagnetic Separation, Staining

Journal: The Journal of Clinical Investigation

Article Title: Liver sinusoidal endothelial cell progenitor cells promote liver regeneration in rats

doi: 10.1172/JCI58789

Figure Lengend Snippet: Resident SPCs (CD133+ fraction of LSECs), LSECs with removal of resident SPCs (fraction of LSECs depleted of CD133+ cells), periportal LSECs (CD45bright), and centrilobular LSECs (CD45– LSECs) were stained for VEGF-receptor 1 (VEGF-R1) and VEGF-R2. The third column shows a merged image. Original magnification, ×100.

Article Snippet: Microbeads from the following kits were used: CD133 Cell Isolation Kit and Anti-FITC MultiSort Kit (Miltenyi Biotec).

Techniques: Staining

Journal: The Journal of Clinical Investigation

Article Title: Liver sinusoidal endothelial cell progenitor cells promote liver regeneration in rats

doi: 10.1172/JCI58789

Figure Lengend Snippet: Two months after neonates were injected with BrdU, LSECs were isolated by elutriation, and the CD133+ fraction was isolated by immunomagnetic separation to obtain resident SPCs and LRCs. (A) CD31 staining of resident SPCs and LRCs. (B) CD45 staining of resident SPCs and LRCs. (C) BrdU+ staining indicates LRCs (white arrows). (D) Differential interference contrast (DIC) demonstrates all cells in the field. (E) The merged image shows CD133+ cells that are CD31+CD45+BrdU+ LRCs (white), indicated by white arrows. Original magnification, ×100.

Article Snippet: Microbeads from the following kits were used: CD133 Cell Isolation Kit and Anti-FITC MultiSort Kit (Miltenyi Biotec).

Techniques: Injection, Isolation, Immunomagnetic Separation, Staining, BrdU Staining

Journal: The Journal of Clinical Investigation

Article Title: Liver sinusoidal endothelial cell progenitor cells promote liver regeneration in rats

doi: 10.1172/JCI58789

Figure Lengend Snippet: LSECs were isolated on day 3 after PHx from wild-type rats that had been transplanted with BM from Lew-Tg(CAG-EGFP)ys rats. (A) Hgf mRNA expression was measured by real-time PCR in GFP+ (BM-derived) and GFP– LSECs. n = 3; ***P < 0.001. (B) HGF protein expression was determined by immunoblot and (C) quantified by densitometry in control LSECs (isolated from nonoperated rats) and in GFP+ and GFP– LSECs isolated on day 3 after PHx (n = 3). *P < 0.05, **P < 0.01 compared with GFP– LSECs after PHx. (D) CD133+ cells from the LSEC fraction were immunostained for PCNA and sorted by FACS for GFP. n = 3; ***P < 0.0001 comparison of the percentage PCNA+ cells in the GFP+ and GFP– fractions.

Article Snippet: Microbeads from the following kits were used: CD133 Cell Isolation Kit and Anti-FITC MultiSort Kit (Miltenyi Biotec).

Techniques: Isolation, Expressing, Real-time Polymerase Chain Reaction, Derivative Assay, Western Blot, Control, Comparison