Journal: Differentiation; research in biological diversity

Article Title: The combination of glycosaminoglycans and fibrous proteins improves cell proliferation and early differentiation of bovine primary skeletal muscle cells.

doi: 10.1016/j.diff.2013.06.006

Figure Lengend Snippet: Fig. 2. Localization and expression of myogenic regulatory transcription factors (MRFs) during proliferation and differentiation. (A) Proliferating cells were fixed with 2% PFA, immunostained with Alexa 488-Phalloidin (green), and rabbit anti-MyoD, followed by DyLight 649-conjugated donkey anti-rabbit (red) before fluorescence microscopy analysis. Nuclei were stained with DAPI (blue). Scale bar 10 mM. (B) Cells differentiated for 3 days were fixed with 2% PFA, immunostained with Alexa 488-Phalloidin (green), and goat anti-Myogenin, followed by DyLight 649-conjugated donkey anti-goat (red) before fluorescence microscopy analysis. Nuclei were stained with DAPI (blue). Arrows indicate Myogenin positive cells. Scale bar 10 mM. (C) Differentiating cells were fixed with 2% PFA, immunostained with Alexa 488-Phalloidin (green) and rabbit anti-Desmin, followed by DyLight 549-conjugated mouse anti-rabbit (red) before fluorescence microscopy analysis. Nuclei were stained with DAPI (blue). Scale bar 10 mM. (D) Left: a representative western blot showing the expression level of MyoD and Desmin after 0 and 3 days of differentiation. Cell lysates were subjected to western blotting using antibodies to MyoD, Desmin, and Tubulin (loading control). Right: quantification of Western blots. Bars show the expression of Desmin and MyoD, relative to tubulin, after 0 and 3 days of differentiation. The graph represents the average of three experiments with SD.

Article Snippet: Alexa 488-conjugated goat anti-mouse and DyLight 549- conjugated mouse anti-rabbit were from Jackson ImmunoResearch Laboratories Inc. (West Grove, PA, USA).

Techniques: Expressing, Microscopy, Staining, Western Blot, Control

Journal: Differentiation; research in biological diversity

Article Title: The combination of glycosaminoglycans and fibrous proteins improves cell proliferation and early differentiation of bovine primary skeletal muscle cells.

doi: 10.1016/j.diff.2013.06.006

Figure Lengend Snippet: Fig. 6. Desmin staining of myotubes on different surface coatings after 1, 3 or 5 days in differentiation medium. Differentiating cells were fixed with 2% PFA and immunostained with rabbit anti-Desmin, followed by DyLight 549-conjugated mouse anti-rabbit (yellow) before fluorescence microscopy analysis. Nuclei were stained with DAPI (blue). Scale bar 20 mM.

Article Snippet: Alexa 488-conjugated goat anti-mouse and DyLight 549- conjugated mouse anti-rabbit were from Jackson ImmunoResearch Laboratories Inc. (West Grove, PA, USA).

Techniques: Staining, Microscopy

Journal: bioRxiv

Article Title: L-DOS47 enhances response to immunotherapy in pancreatic cancer tumor

doi: 10.1101/2023.08.28.555194

Figure Lengend Snippet: A . Percentage of CEACAM6-expressing cells analyzed by flow cytometry in KPC961 parental and KPC961 clone 1B6 cells. B . Ratio of extracellular acidification (ECAR) and oxygen consumption rate (OCR) profiles in KPC961 parental and clone 1B6 cells; C . Lactate production in KPC961 parental and clone 1B6 cells. D . Tumor growth as a function of inoculum size. KPC961-CEACAM6 cells were inoculated into the pancreas of B6.129 mice at 100000, 50000, and 20000 cells, and tumor volumes were measured by ultrasound. Ascites fluid was observed in two mice inoculated with 100,000 cells on days 24 and 29, two mice inoculated with 50,000 cells at day 29 and one mouse inoculated with 20,000 cells at day 27. E . Expression of CEACAM6, PD-L1 and H2Kb in KPC961-1B6 cells of inoculated orthotopic tumors by flow cytometry.

Article Snippet: Cells were labeled with the following antibodies:(CEACAM6-FITC at 10 μg/mL, Sino Biological 10823-R408R; H2Kb-Pac Blue at 0.5 mg/mL, BioLegend 116514; CD45-BV605 at 0.2 mg/mL, BioLegend 103155; and PD-L1 – PE at 0.2 mg/mL, Invitrogen 12-5982-82) in FACS buffer for 20 minutes at 4°C in the dark.

Techniques: Expressing, Flow Cytometry