Journal: PLoS Pathogens

Article Title: Receptor-Targeted Nipah Virus Glycoproteins Improve Cell-Type Selective Gene Delivery and Reveal a Preference for Membrane-Proximal Cell Attachment

doi: 10.1371/journal.ppat.1005641

Figure Lengend Snippet: ( A ) Cell entry is EpCAM dependent but independent of ephrin-B2. Representative flow cytometry plots out of three independent experiments of CHO-K1, CHO-EpCAM, CHO-ephrin-B2 cells, and of a mixed culture composed of CHO-EpCAM and CHO-ephrin-B2 (1:1 ratio) monitored 72 h after transduction with NiVmut EpCAM -LV, NiVwt-LV or VSV-LV (MOI of 1). EpCAM expression was detected by an APC-coupled human EpCAM specific antibody. ( B ) To ascertain stability of transduction with the EpCAM-targeted vector, CHO-EpCAM cells were cultivated for further 30 days after transduction with the indicated MOIs. The percentage of GFP-positive cells was determined by flow cytometry at the indicated time points. One representative out of three independent experiments is shown.

Article Snippet: Human EpCAM was detected by an Allophycocyanin (APC) labeled mouse anti-EpCAM antibody (clone HEA-125, Miltenyi Biotec, Bergisch Gladbach, Germany, dilution 1:100).

Techniques: Flow Cytometry, Transduction, Expressing, Plasmid Preparation

Journal: Genes & Cancer

Article Title: CD133-positive cancer stem cells from Colo205 human colon adenocarcinoma cell line show resistance to chemotherapy and display a specific metabolomic profile

doi:

Figure Lengend Snippet: A-B. Cytometry analyses of Nestin, CK20, CD133, Oct3/4, CD44 and CD44v9 on 10% FBS Colo205 growing cells (control) and serum-free growing cultures (week 1 to week 5).

Article Snippet: The purity of sorted cells was evaluated by flow cytometry using FACSCalibur (BD Biosciences) after labeling with anti-human CD133/2 or CD44 antibody (Miltenyi Biotec).

Techniques: Cytometry, Control

Journal: Genes & Cancer

Article Title: CD133-positive cancer stem cells from Colo205 human colon adenocarcinoma cell line show resistance to chemotherapy and display a specific metabolomic profile

doi:

Figure Lengend Snippet: A. Tumorsphere evaluation of CD133+ and CD133- sorted cells. B. Invasiveness of different cell fractions measured using a collagen-based invasion kit (relative DO measured at 560 nm). C. Survival assay after chemotherapy treatment with cisplatin and 5-FU. D. Metabolite quantification after CE-TOF-MS experiments. Only metabolites of interest for Colo205 cells cultured in 10% FBS (control; green bar), CD133+ sorted cells (red bar) and CD44+ sorted cells (yellow bar) are reported here.

Article Snippet: The purity of sorted cells was evaluated by flow cytometry using FACSCalibur (BD Biosciences) after labeling with anti-human CD133/2 or CD44 antibody (Miltenyi Biotec).

Techniques: Clonogenic Cell Survival Assay, Cell Culture, Control

Journal: International journal of molecular medicine

Article Title: Autophagy is essential for the endothelial differentiation of breast cancer stem‑like cells.

doi: 10.3892/ijmm.2019.4399

Figure Lengend Snippet: Figure 1. Flow cytometry and microsphere culture of BCSLCs. (A) CD24‑/low cells were isolated from MCF‑7 using immunomagnetic beads, and the expres- sion of CD24 in MCF‑7 and isolated CD24‑/low cells were assessed using flow cytometry. (a) Isotype control for (b); (b) MCF‑7 cells; (c) isotype control for (d); (d) isolated CD24‑/low cells. (B) CD44+ cells were further isolated from CD24‑/low cells and the expression of CD44 was assessed using flow cytometry. (a) Isotype control for (b); (b) CD44+ cells. (C) Expression of CD24 and CD44 in MCF‑7, BCSLCs and BCSLCs after eight passages. (a) isotype control for (b); (b) MCF‑7 cells; (c) CSLCs; (d) isotype control for (e); (e) BCSLCs after eight passages. (D) The isolated BCCSLCs were cultured in microspheres for 0 and 64 h in stem cell culture medium. BCSLC, breast cancer stem‑like cell; PE, phycoerythrin.

Article Snippet: The following primary antibodies were used: Fluorescein isothiocyanate (FITc)-conjugated anti-human cd44 (cat. no. 130-113-903; Miltenyi Biotec GmbH), phycoerythrin (PE)-conjugated anti-human cd24 (cat. no. 130-098-861; Miltenyi Biotec GmbH) PE-conjugated anti-human cd31 (cat. no. 130-110-807; Miltenyi Biotec GmbH) and FITc-conjugated anti-human cd105 (cat. no. 130-098-778; Miltenyi Biotec GmbH).

Techniques: Flow Cytometry, Isolation, Control, Expressing, Cell Culture, Stem Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Cellular Mechanisms of the Anti-Arrhythmic Effect of Cardiac PDE2 Overexpression

doi: 10.3390/ijms22094816

Figure Lengend Snippet: Ca 2+ /calmodulin-dependent kinase II (CaMKII)-mediated increase in I NaL and cellular arrhythmogenic events upon β-AR stimulation is abolished in PDE2 OE. Late sodium current (I NaL ) was measured in isolated ventricular cardiomyocytes from PDE2 OE (squares) and WT (circles) mice at −35 mV (1 s) under basal conditions (black symbols) and after 10 min stimulation with ISO (100 nM, orange), ISO (100 nM) and AIP (1 µM, blue), ISO (100 nM) and PKI (5 µM, green), 8-CPT (10 µM, red), 8-CPT (10 µM) and AIP (1 µM, dark blue). ( A , B ) Representative current traces and ( C , D ) quantification of I NaL estimated by integrating I Na between 100 and 500 ms and normalized to the membrane capacitance. *** p < 0.001 vs. control (Ctrl), (N = animals/genotype, 6 ≤ n ≤ 17 cells per group). ( E , F ) Isolated ventricular cardiomyocytes were subjected to an arrhythmia provocation protocol under basal conditions (black), after 10 min stimulation with ISO (100 nM, orange) or ISO (100 nM) and AIP (1 µM, blue). Arrhythmogenic events (DADs, EADs, and sAPs) were quantified. *** p < 0.001; 8≤ n ≤ 13, (N = 5 animals/group).

Article Snippet: For protein quantification the following specific primary antibodies were used: Epac1 (cell signaling, 5D3, #4155), Epac2 (cell signaling, #4156S), CaMKIIδ total (R&D Systems, MAB4176), CaMKII pThr286 PKAIIα (Thermo, MA1-047), PKA-C (BD Bioscience, #610981), PKA IIα reg (C20) (Santa-Cruz, sc-908), PKCα (H-7) (Santa-Cruz, sc-8393), NCX1 (Swant, π 11-13), NKA (cell signaling #3010), PLM (Abcam, ab76579), EEF2 (Abcam, ab40812), GAPDH (Santa-Cruz, sc-365062).

Techniques: Isolation, Membrane, Control

Journal: International Journal of Molecular Sciences

Article Title: Cellular Mechanisms of the Anti-Arrhythmic Effect of Cardiac PDE2 Overexpression

doi: 10.3390/ijms22094816

Figure Lengend Snippet: Basal expression and phosphorylation of CaMKII is affected by PDE2 overexpression. ( A ) Representative immunoblots and ( B ) quantification of protein kinase expression normalized to GAPDH, CaMKII phosphorylation at Threonine 286 normalized to total CaMKIIδ in lysates of isolated cardiomyocytes from PDE2 OE (orange) and WT (black) mice. (N = 6 animals/genotype. * p < 0.05, *** p < 0.001 vs. WT).

Article Snippet: For protein quantification the following specific primary antibodies were used: Epac1 (cell signaling, 5D3, #4155), Epac2 (cell signaling, #4156S), CaMKIIδ total (R&D Systems, MAB4176), CaMKII pThr286 PKAIIα (Thermo, MA1-047), PKA-C (BD Bioscience, #610981), PKA IIα reg (C20) (Santa-Cruz, sc-908), PKCα (H-7) (Santa-Cruz, sc-8393), NCX1 (Swant, π 11-13), NKA (cell signaling #3010), PLM (Abcam, ab76579), EEF2 (Abcam, ab40812), GAPDH (Santa-Cruz, sc-365062).

Techniques: Expressing, Phospho-proteomics, Over Expression, Western Blot, Isolation