Journal: World Journal of Stem Cells

Article Title: Patient-derived induced pluripotent stem cells with a MERTK mutation exhibit cell junction abnormalities and aberrant cellular differentiation potential

doi: 10.4252/wjsc.v16.i5.512

Figure Lengend Snippet: Generation and identification of the human induced pluripotent stem cells derived from the retinitis pigmentosa patient. A: Timeline of human induced pluripotent stem cell generation; B: Imaging by phase-contrast microscopy. Scale bar = 200 μm; C: Karyotype analysis of the healthy control (left) and retinitis pigmentosa patient (right); D: Flow cytometry of pluripotency markers SSEA-4 and TRA-1-81; E and F: Immunostaining of pluripotency markers OCT4, NANOG, SOX2, and SSEA4 of the healthy control and retinitis pigmentosa patient. Scale bar = 20 μm; G and H: In vitro differentiation of control (G) and patient (H) induced pluripotent stem cells into three germ layers, endoderm (AFP+), mesoderm (α-SMA+) and ectoderm (GFAP+). Scale bar = 20 μm. PB: Peripheral blood; PBMC: Peripheral blood mononuclear cell; iPSC: Induced pluripotent stem cell.

Article Snippet: Following primary antibodies were used: OCT4 (Cat. #ab18976; Abcam), SOX2 (Cat. #sc-365823; Santa Cruz), NANOG (Cat. #ab80892; Abcam), SSEA4 (Cat. #ab16287; Abcam), GFAP (Cat. #HPA056030; Sigma), α-SMA (Cat. #A5228; Sigma), AFP (Cat. #MAB1368; R&D System).

Techniques: Derivative Assay, Imaging, Microscopy, Control, Flow Cytometry, Immunostaining, In Vitro

Journal: Frontiers in Physiology

Article Title: Pathogenesis of Molar Hypomineralisation: Hypomineralised 6-Year Molars Contain Traces of Fetal Serum Albumin

doi: 10.3389/fphys.2020.00619

Figure Lengend Snippet: A molecular approach for timestamping exposures of serum albumin to dental enamel. (A) Schematic showing three hypothetical stages where albumin could become incorporated in enamel on a 6-year molar ( yellow spot ), as considered in this study. These albumin exposures are designated developmental (0–3 years), eruptive (6 years), or artefactual during subsequent extraction, as indicated. (B) Serum levels of serum albumin (ALB; dark blue ) and alpha-fetoprotein (AFP; cyan ) in healthy individuals aged from 3-months postconception through to 1-year old, as indicated. Data were collated and averaged from 9 population studies as outlined in Methods and . It can be seen that at birth ( dotted line ), ALB is about 600-fold more concentrated than AFP. (C) The adult/fetal isoform ratio for serum albumin ( black line ), as derived from (B) , rises quasi-linearly through to 6-months of age ( cream box ). Sensitivity limits for the AFP immunoblot assay reached the early-post-natal period ( cyan box ). Note the isoform ratio, which spans 6-orders of magnitude (cf. log scale), offers strong resolution of age in early infancy.

Article Snippet: Human albumin (Sigma), alpha-fetoprotein (Lee Biosolutions), recombinant alpha-fetoprotein and albumin (both tagged with glutathione S-transferase; from Abnova), and rabbit polyclonal antibodies to human albumin (PAB10220 from Abnova), alpha-fetoprotein peptide (PAB12795 from Abnova) and whole alpha-fetoprotein (H00000174-D01 from Abnova), were obtained commercially.

Techniques: Extraction, Derivative Assay, Cream, Western Blot

Journal: Cell reports

Article Title: Critical roles of translation initiation and RNA uridylation in endogenous retroviral expression and neural differentiation in pluripotent stem cells

doi: 10.1016/j.celrep.2020.107715

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Antibodies and dilution rate were as follows; mouse monoclonal anti-OCT3/4 (1:200, sc-5279, Santa Cruz Biotechnology), rabbit polyclonal anti-SOX2 (1:100, ab97959, Abcam), rabbit polyclonal anti-NANOG (1:100, ab21624, Abcam), mouse monoclonal anti-human Nuclei (1:1000, MAB4383, EMD Millipore), rabbit polyclonal anti-AFP (1:200, A0008, DAKO), mouse monoclonal anti-AFP (1:200, MAB1368, R&D Systems), mouse monoclonal anti-SMA (1:100, M0851, DAKO), mouse monoclonal anti-βIII-TUBULIN (1:1000, MAB1637, EMD Millipore), rabbit polyclonal anti-PAX6 (1:1000, 901301, BioLegend), rabbit polyclonal anti-KLF4 (1:100, sc-20691, Santa Cruz Biotechnology), goat polyclonal anti-TFCP2L1 (1:100, AF5276, R&D Systems), rabbit polyclonal anti-TFE3 (1:100, HPA023881, Sigma-Aldrich), Alexa 488-conjugated donkey anti-mouse IgG (1:500, A-21202, Thermo Fisher Scientific), Alexa 555-conjugated donkey anti-rabbit IgG (1:500, A-31572, Thermo Fisher Scientific), Alexa 647-conjugated donkey anti-mouse IgG (1:500, A-31571, Thermo Fisher Scientific), Alexa 488-conjugated anti-rabbit IgG (1:500, A-21206, Thermo Fisher Scientific), Alexa 555-conjugated donkey anti-rabbit IgG (1:500, A-31572, Thermo Fisher Scientific), Alexa 647-conjugated donkey anti-rabbit IgG (1:500, A-31573, Thermo Fisher Scientific) and Alexa 488-conjugated donkey anti-goat IgG (1:500, A-11055, Thermo Fisher Scientific).

Techniques: Recombinant, Protease Inhibitor, Modification, Knock-Out, Transfection, Blocking Assay, Western Blot, Immunocytochemistry, Lysis, Purification, Clone Assay, SYBR Green Assay, Reporter Assay, TaqMan Assay, Expressing, Microarray, shRNA, Plasmid Preparation, Software

Journal: Proteomics

Article Title: Development and application of a novel recombinant Aleuria aurantia lectin with enhanced core fucose binding for identification of glycoprotein biomarker of hepatocellular carcinoma

doi: 10.1002/pmic.201600064

Figure Lengend Snippet: (A) SDS-PAGE and Coomassie Blue detection of purchased AAL (Vector Lab) or recombinant AAL (rAAL) with 10×His tag, rAAL with 10×His and N224Q mutation. (B) SDS-PAGE and Coomassie Blue staining of glycoprotein antigens. From left to right, human IgG (IgG), α-1,3 fucose (F-BSA), galactosylated BSA (G-BSA), alpha fetoprotein (AFP). (C) Lectin blotting of samples in Panel B with rAAL. Lectin blotting of samples in Panel B with the N224Q lectin (E) Lectin blotting of samples in Panel B with PhoSL. In all cases lectins were biotinylated and visualized using LI-COR Odyssey Infrared scanner under the same intensity (5.0).

Article Snippet: Human cord serum alpha-fetoprotein (AFP) was purchased from Lee Biosolutions, Inc. (Maryland Heights, MO, USA).

Techniques: SDS Page, Plasmid Preparation, Recombinant, Mutagenesis, Staining