Journal: Nature neuroscience

Article Title: Therapeutically viable generation of neurons with antisense oligonucleotide suppression of PTB.

doi: 10.1038/s41593-021-00864-y

Figure Lengend Snippet: Fig. 3 | Transition of GFAP:tdTomato+ cells into new neurons results in no depletion in the total GFAP cell number. a, Images of newly induced neurons (iNeurons) expressing NeuN (green) and tdTomato (red) (NeuN visualized by IF and tdTomato by direct fluorescence); blue, DAPI staining for DNA; white, GFAP visualized by IF; the experiment was reproduced four times, independently, with similar results. b, Total number of tdTomato+NeuN− cells (left) or tdTomato+NeuN+ cells (right), either with or without GFAP labeling, per dentate gyrus, 2 months after ICV injection of PTB-ASO. Data are presented as mean ± s.e.m. (left: tdTomato/NeuN: 176.8 ± 34.92; tdTomato/NeuN/GFAP mean: 167 ± 34.32; right: tdTomato/NeuN mean: 28.88 ± 4.87; tdTomato/NeuN/GFAP mean: 0.87 ± 0.12; n = 4 biological repeats; two-tailed t-test **P = 0.0012). c, Representative images of human organoids 1 month after PTB-ASO or control-ASO application in the growth media GFAP (green) and Nestin (red) (both visualized by IF); blue, DAPI staining for DNA; the experiment was reproduced three times, independently, with similar results. d, Representative images of a dentate gyrus taken 2 months after ICV injection of control or PTB-ASO2 into 3-month-old and 1-year-old mice; white, GFAP visualized by IF; the experiment was reproduced three times for control and aged mice and four times for 5-month-old mice, independently, with similar results. e, Total GFAP-positive cells per dentate gyrus area of 5-month- and 1.2-year-old PTB-ASO- or control-injected mice. Data are presented as mean ± s.e.m. (control mean: 1,074 ± 83.55; 5 months PTB-ASO mean: 1,006 ± 82.45; 1.2 year-old PTB-ASO mean: 1,214 ± 104.8; n = 3, n = 4, n = 3 biological repeats).

Article Snippet: Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Human research participants Clinical data Dual use research of concern Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Rabbit-PTBP1 (Abclonal; Cat# - A6107; IF concentration - 1:1000); Mouse-GFAP (Dako; Cat# - M0761; IF concentration - 1:500); Mouse-NeuN (Millipore; Cat# - MAB377; IF concentration - 1:100); Rabbit-pan-ASO (Ionis pharmaceutical, homemade antibody; Cat# -13545; IF concentration - 1:200); Chicken-MAP2 (Abcam; Cat# - ab5392; IF concentration - 1:1000); Mouse-Nestin (abclonal; Cat# - AB22035; IF concentration - 1:200); Rabbit-GFAP (Novus Biologicals; Cat# - NB300-141; IF concentration - 1:1000); Mouse-TubIII (Tuj1) (Millipore Sigma; Cat# - T8660: IF concentration - 1:500); Goat-DCX (Santa Cruz Biotechnology; Cat# - Sc-8066; IF concentration - 1:500).

Techniques: Expressing, Fluorescence, Staining, Labeling, Injection, Two Tailed Test, Control

Journal: International Journal of Molecular Sciences

Article Title: Cold Atmospheric Plasma Does Not Affect Stellate Cells Phenotype in Pancreatic Cancer Tissue in Ovo

doi: 10.3390/ijms23041954

Figure Lengend Snippet: CAP does not alter the activation profile of RLT-PSC cells in ovo. Sections of RLT-PSC and co-cultured RLT-PSC + Mia PaCa-2 from in ovo tissue were stained for GFAP, vimentin (Vim), and ACTA-2. ( a – c , g – i ) Mean fluorescence intensity (MFI) expressed as arbitrary units (a.u.); ( d – f , j – l ) number of cells positive for the corresponding markers. Anti-CD44 antibodies (PDAC cells) were used as a counterstain to discriminate Mia PaCa-2 cells from RLT-PSC. UT = untreated control. n ≥ 4 tissue per condition; each dot represents one tissue. Dashed lines = median; dotted lines = 25% and 75% quartiles; * = p ≤ 0.05.

Article Snippet: Primary antibodies used: GFAP (HPA056030, Atlas Antibodies, 1/200, Bromma, Switzerland); ACTA-2 (M0851, Dako, 1/200); Vimentin (Abcam ab92547, 1/500, Cambridge, UK); CD44 (3570S, Cell Signaling Technologies, 1/400, Danvers, MA, USA); MMP2 (40994, Cell Signaling Technologies, 1/400, Danvers, MA, USA); MMP9 (13667, Cell Signaling Technologies, 1/100, Danvers, MA, USA).

Techniques: Activation Assay, In Ovo, Cell Culture, Staining, Fluorescence, Control

Journal: International Journal of Molecular Sciences

Article Title: Cold Atmospheric Plasma Does Not Affect Stellate Cells Phenotype in Pancreatic Cancer Tissue in Ovo

doi: 10.3390/ijms23041954

Figure Lengend Snippet: List of primers used for quantitative real-time PCR.

Article Snippet: Primary antibodies used: GFAP (HPA056030, Atlas Antibodies, 1/200, Bromma, Switzerland); ACTA-2 (M0851, Dako, 1/200); Vimentin (Abcam ab92547, 1/500, Cambridge, UK); CD44 (3570S, Cell Signaling Technologies, 1/400, Danvers, MA, USA); MMP2 (40994, Cell Signaling Technologies, 1/400, Danvers, MA, USA); MMP9 (13667, Cell Signaling Technologies, 1/100, Danvers, MA, USA).

Techniques:

Journal: Gene therapy

Article Title: Targeted inhibition of platelet-derived growth factor receptor-beta subunit in hepatic stellate cells ameliorates hepatic fibrosis in rats.

doi: 10.1038/gt.2008.93

Figure Lengend Snippet: Figure 3 Localization of LacZ driven by CMV promoter and GFAP promoter in normal rats, acute liver injury rats and chronic liver injury rats. (a) Representative graphs of b-galactosidase (b-gal) immunofluorescence showed the distribution of b-gal-positive cells in normal rats, acute liver injury rats and chronic liver injury rats treated with pCMV-shRNA-LacZ or pGfa-shRNA-LacZ. Magnification of 20. The scale bar represents 80 mm. (b) Representative graphs of a-SMA and GFAP immunofluorescence showed that in the chronic injured liver, a-SMA and GFAP proteins distributed mainly around the portal area and the hyperplastic bile duct, whereas in the acute injured liver, both the proteins revealed a much more diffuse distribution in the hepatic lobule. In addition, the GFAP-positive cells were more than the a-SMA- positive cells both in the acute injured liver and in the chronic injured liver. Magnification of 20. The scale bar represents 80 mm. (c) Representative graphs of b-gal and a-SMA double-staining revealed that b-gal protein in pCMV-shRNA-LacZ-treated livers could be expressed in hepatocytes (blue arrow) and activated HSCs stained by a-SMA (white arrow), magnification of 63. The scale bar represents 40 mm. (d) Representative graphs of b-gal and GFAP double-staining showed that b-gal-positive cells were all GFAP-positive HSCs (pink arrow) in pGfa-shRNA-LacZ-treated livers, magnification of 63. The scale bar represents 40 mm. (e) Representative graphs of b-gal and a-SMA double-staining showed that some b-gal protein was expressed in activated HSCs stained by a-SMA (white arrow) in pGfa-shRNA- LacZ-treated livers, magnification of 63. The scale bar represents 40 mm. CMV, cytomegalovirus; GFAP, glial fibrillary acidic protein; HSCs, hepatic stellate cells; PDGFR-b, platelet-derived growth factor receptor-b subunit; a-SMA, a-smooth muscle actin; shRNA, short hairpin RNA; RT-PCR, reverse transcriptional-PCR.

Article Snippet: Frozen liver sections (8 mm in thickness) were incubated with mouse anti-b-galactosidase monoclonal antibody (Promega, Madison, WI, USA), rabbit anti-GFAP polyclonal antibody (Boster, Wuhan, China) or rabbit anti-a-SMA polyclonal antibody (Bios, Beijing, China) for 20 h at 4 1C, then washed in phosphate-buffered saline and incubated for 30 min with Cy5-labeled anti-rabbit IgG antiserum and FITC-labeled anti-mouse IgG antiserum (Jackson, West Grove, PA, USA) at 1:100 dilution in phosphate-buffered saline.

Techniques: shRNA, Double Staining, Staining, Derivative Assay, Reverse Transcription Polymerase Chain Reaction