Journal: STAR Protocols

Article Title: Preparation of mouse pancreatic tumor for single-cell RNA sequencing and analysis of the data

doi: 10.1016/j.xpro.2021.100989

Figure Lengend Snippet:

Article Snippet: Dead Cell Removal Kit , Miltenyi Biotec , , 130-090-101.

Techniques: Recombinant, Saline, Modification, Staining, Red Blood Cell Lysis, Multiplex Assay, Sequencing, Generated, Software, Sterility

Journal: STAR Protocols

Article Title: Preparation of mouse pancreatic tumor for single-cell RNA sequencing and analysis of the data

doi: 10.1016/j.xpro.2021.100989

Figure Lengend Snippet: 1 × Binding Buffer (Dead cell removal kit)

Article Snippet: Dead Cell Removal Kit , Miltenyi Biotec , , 130-090-101.

Techniques: Binding Assay, Concentration Assay, Sterility

Journal: International Journal of Molecular Sciences

Article Title: Development of a High-Efficacy Reprogramming Method for Generating Human Induced Pluripotent Stem (iPS) Cells from Pathologic and Senescent Somatic Cells

doi: 10.3390/ijms21186764

Figure Lengend Snippet: TRA-1-60 and alkaline phosphatase staining of iPS cell colonies induced from the dermal tissue of three elderly patients. ( a ) TRA-1 immunofluorescent staining of iPS cell colonies. Immunofluorescent staining with anti-TRA1-60 antibody was performed on D1 (top), D2 (middle), and D3 (bottom) cells subjected to protocol 1 (left) or protocol 2 (right) for iPS cell induction at day 35 post-transfection. An integration of 81 continuous fields of the view captured with the Keyence BZ-X710 microscope is shown (observed area: 23.915 mm × 17.936 mm). BF, bright field. Arrows indicate TRA1-60-positive iPS cell colonies. Scale bars: 3.0 mm. ( b ) Alkaline phosphatase staining of iPS cell colonies. Staining with alkaline phosphatase was performed on D1 (top), D2 (middle), and D3 (bottom) cells subjected to protocol 1 (left) or protocol 2 (right) for iPS cell induction at day 36 post-transfection. Arrows indicate alkaline phosphatase-positive iPS cell colonies. ( c ) The number of iPS cell colonies stained by alkaline phosphatase was quantitively analyzed in all fibroblasts. The left side is protocol 1 and the right side is protocol 2. Values shown are the mean ± SEM (* p < 0.05, n = 3).

Article Snippet: An Anti-TRA-1-60 antibody, anti-SSEA4 antibody, and alkaline phosphatase stain kit (AP Staining Kit, Red-Color) was purchased from Funakoshi ® (Tokyo, Japan).

Techniques: Staining, Transfection, Microscopy

Journal: bioRxiv

Article Title: IL-11 neutralising therapies target hepatic stellate cell-induced liver inflammation and fibrosis in NASH

doi: 10.1101/470062

Figure Lengend Snippet: a , IL-11 is upregulated in hepatic stellate cells (HSCs) stimulated with TGFβ1 (n=3). b , IL-11 protein is secreted from HSCs stimulated with TGFβ1 (ELISA, n=3). c , Human precision cut liver slices were stimulated with TGFβ1 and IL-11 protein was measured in supernatant (ELISA, n=3). d , IL6R, gp130, and IL11RA expression in HSCs (TPM, transcripts per million). e, f , Representative fluorescence images (scale bars, 200 µm) of HSCs and automated fluorescence quantification for ( e ) ACTA2 +ve cells and ( f ) Collagen I immunostaining following incubation without stimulus (-), with TGFβ1, PDGF, or IL-11. g , Collagen secretion supernatants of HSC stimulated with TGFβ1, PDGF, or IL-11 (Sirius red assay. n≥7). h , Dose-dependent matrigel invasion of HSCs induced by IL-11(n=3). i , Hyper IL-11 induces IL-11 protein secretion from HSCs (ELISA, n=3). a-c, e-g, i , TGFβ1 (5 ng ml -1 ), Hyper IL-11 (0.2 ng ml -1 ), PDGF (20 ng ml -1 ), IL-11 (5 ng ml -1 ); 24 h stimulation; h , 48 h stimulation. j , Schematic of mice receiving daily subcutaneous injection of either saline (control) or rmIl-11 (100 µg kg -1 ). k , Relative liver hydroxyproline content, l , mRNA expression of pro-fibrotic markers, and m , serum ALT levels (k, l , control, n=7; rmIl-11, n=10; m , control n=8; rmIl-11, n=11). a, b, g, h, i, k, m Data are represented as mean ±s.d; c-f, l , Box-and-whisker plots show median (middle line), 25th–75th percentiles (box) and min-max percentiles (whiskers). a-d, i, k-m , Two-tailed Student’s t -test; e-h , two-tailed Dunnett’s test. FC: fold change; I/A: intensity/area.

Article Snippet: Total secreted collagen in the cell culture supernatant was quantified using Sirius red collagen detection kit (9062, Chondrex).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Fluorescence, Immunostaining, Incubation, Injection, Whisker Assay, Two Tailed Test