Journal: Animal Nutrition

Article Title: Arginine promotes myogenic differentiation and myotube formation through the elevation of cytoplasmic calcium concentration

doi: 10.1016/j.aninu.2021.05.010

Figure Lengend Snippet: NIS blocked arginine-promoted myogenic differentiation of C2C12 myoblasts. C2C12 cells at 80%-90% confluence were cultured in differentiation medium and treated with 1.2 mmol/L arginine in the absence or presence of NIS (10 μmol/L) for 4 (A, B) or 2 d (C, D). (A) Immunostaining of C2C12 myotubes using antibody against myosin. (B) Myotube area is calculated as described in methods. (C, D) Relative mRNA expression levels of MyoD1 , MyoG , Cav1.1 , RyR1 and Stim1 normalized to GAPDH mRNA, measured by real-time quantitative PCR in triplicate. Values are means ± SEM ( n = 3). ∗ P < 0.05. ∗∗ P < 0.01. Scale bars, 100 μm. CON, control; Arg, 1.2 mmol/L arginine; NIS, 10 μmol/L nisoldipine.

Article Snippet: Dantrolene (DAN) and nisoldipine (NIS) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Cell Culture, Immunostaining, Expressing, Real-time Polymerase Chain Reaction, Control

Journal: Neuroscience

Article Title: Effects of crenolanib, a non-selective inhibitor of PDGFR, in a mouse model of transient middle cerebral artery occlusion

doi: 10.1016/j.neuroscience.2017.09.025

Figure Lengend Snippet: (A–C) Immunofluorescence staining of proliferative vessels (CD31+/BrdU+) in the peri-infarct region on day 7 after MCAO. CD31: green, BrdU: red. Images are shown at 200× magnification; scale bar=50 μm. (D) Quantification showed that crenolanib treatment on days 1–3 significantly decreased the number of proliferative vessels in the peri-infarct area on day 7 after stroke. *p<0.05 vs. MCAO+vehicle group; n=8/group. (E) ELISA analysis of ischemic border on day 7 after MCAO showed that crenolanib treatment on days 1–3 decreased the average level of VEGF in MCAO mice, but the change was not statistically significant. n=8/group. (F–G) Immunofluorescence staining of SDF-1 and MCP-1 in the peri-infarct region on day 7 after MCAO. The white lines show the brain boundaries. Images are shown at 100× magnification; scale bar=25 μm. (H–I) ELISA analysis of SDF-1 and MCP-1 on day 7 after MCAO showed that levels of SDF-1 and MCP-1 in the ischemic border were lower in MCAO mice treated with crenolanib (on days 1–3) than in MCAO mice treated with vehicle. *p<0.05 vs. MCAO+vehicle group; n=8/group.

Article Snippet: Based on our established protocol ( Zhao et al., 2015 , Han et al., 2016 , Wang et al., 2017 ), brain sections (20 μm) were processed with Nissl staining to quantify infarct volume or stained with antibodies against PDGFRβ (1:300, Abcam, Cambridge, MA, USA), GFAP (1:300, Santa Cruz Biotechnology, Dallas, TX, USA), doublecortin (DCX; 1:250, Santa Cruz Biotechnology), cleaved-caspase 3 (cCasp3; 1:500, Millipore, Billerica, MA, USA), CD31 (1:100, Abcam), BrdU (1:250, Abcam), Lectin fluorescein lycopersicon esculentum (tomato) (Lectin, 1:1000; Vector Laboratories, Burlingame, CA), SDF-1 (1:50, Proteintech, Sanying Biotechnology, Wuhan, China), MCP-1 (Proteintech), CD68 (1:500, Santa Cruz Biotechnology), occludin (1:250, Proteintech), and albumin (1:500, Santa Cruz Biotechnology).

Techniques: Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay