Journal: The Journal of Neuroscience

Article Title: Novel Regulatory Mechanisms for the SoxC Transcriptional Network Required for Visual Pathway Development

doi: 10.1523/JNEUROSCI.3430-13.2017

Figure Lengend Snippet: Sox4 promotes human stem cell differentiation into functional RGCs. A–D, Human iPSCs (A, arrows) cultured on eradicated mouse embryonic fibroblast (MEF) feeder layers (A, asterisks) expressed embryonic stem cell markers Oct4 (B) and SSEA-4 (C), but not the neuronal marker βIII-tubulin (D). E, Embryoid bodies dissociated into ≤100 μm aggregates and single cells and plated on PDL and laminin for 24 h showed RPC marker coexpression of Pax6 and Rx. F, During 5 d of differentiation, cells were infected with eGFP (control), Math5-RFP, SOX4-2A-eGFP, or both (example shown in F). G–I, After 5 d, overexpression of SOX4-GFP (G) potentiated RGC morphologic differentiation including long neurites (arrow) with growth cones (arrowhead), and (H and I) increased expression of the RGC marker Brn3. Math5 and Sox4 co-overexpression in human iPSCs together synergized in potentiating RGC-like differentiation (*p < 0.05 from control; **p < 0.01 from control, Math5-alone, and Sox4-alone conditions; N = 3, ANOVA with post hoc Dunnett's test, mean ± SEM shown). J, K, RGC-like progeny generated (J) single and (K) multiple burst action potentials with properties similar to those of purified primary rodent RGCs. Scale bars, 30 μm.

Article Snippet: Commercially available iPSCs (System Biosciences) derived from human foreskin fibroblasts were cultured (1) on irradiated mouse embryonic fibroblast feeder layers (GlobalStem) and (2) in feeder-free conditions on BD Matrigel in mTeSR1 medium (Stem Cell Technologies) supplemented with 5 μ m Y-27632 ROCK inhibitor (Stemgent) and 1 μ m Thiazovivin (Stemgent). iPS colonies were dissociated to single-cell suspension in ultralow attachment plates (Corning).

Techniques: Cell Differentiation, Functional Assay, Cell Culture, Marker, Infection, Over Expression, Expressing, Generated, Purification

Journal: Nature Communications

Article Title: Pancreatic tumor eradication via selective Pin1 inhibition in cancer-associated fibroblasts and T lymphocytes engagement

doi: 10.1038/s41467-022-31928-7

Figure Lengend Snippet: a Establishment and validation of CAFs via high expressions of both FAP-α and α-SMA. b Cellular uptake of Cy5-labeled formulations on Pan02 cells, NIH-3T3 cells, and CAFs, measured by flow cytometry ( n = 3 biologically independent experiments; error bars, mean ± SD). c Cellular uptake of Cy5-labeled antiCAFs-DMS on cells, imaged by fluorescent microscopy. Scale bars are 20 μm. Three independent repetitions got similar results. d Cellular uptake of Cy5-labeled antiCAFs-DMS on cells, quantified by flow cytometry ( n = 6 biologically independent samples; error bars, mean ± SD). Cytotoxicity on Pan02 cells ( e ) and CAFs ( f ) ( n = 3 biologically independent experiments; error bars, mean ± SD). g Western blotting detection of Pin1 and relative proteins from Pan02 cells after treating with PBS, AG17724 (0.5 μM), or antiCAFs-DMS (corresponds to 0.5 μM of AG17724) for 24 hours. h Quantitative analysis of the western blotting results from Pan02 cells (Three independent repetitions got similar results.). i Western blotting detection of Pin1 and relative proteins from CAFs treated with PBS, AG17724, or antiCAFs-DMS after treating with PBS, AG17724 (0.5 μM) or antiCAFs-DMS (corresponds to 0.5 μM of AG17724) for 24 hours. j Quantitative analysis of the western blotting results from CAFs (Three independent repetitions got similar results.). One-way analysis of variance followed by Turkey posttests. Source data are provided as a Source Data file.

Article Snippet: Anti-FAP-α antibody (MAB9727-100, 0.25 μg/mL as working concentration.) and anti-phospho-CDC2/CDK1 (Y15) antibody (AF888-SP, 0.2 μg/mL as working concentration.) were purchased from R&D SYSTEMS.

Techniques: Biomarker Discovery, Labeling, Flow Cytometry, Microscopy, Western Blot