Journal: ASN NEURO

Article Title: Diverse Responses of Oligodendrocytes to Different FGF-Family Members: Uncoupling Structure-Function Relationship Within FGF Subfamilies

doi: 10.1080/17590914.2024.2371163

Figure Lengend Snippet: Differential effects of FGFs on mature oligodendrocyte. Mature OLs were grown in the absence (control) or presence of FGF1, FGF2, FGF4, FGF5, FGF6, FGF9, FGF16, or FGF20 for 2 d, and analyzed for the expression of myelin proteins MOG, FGFR2, BrdU incorporation and p21cip1 expression. a,b. Quantification of the levels of MOG and FGFR2 by immunoblotting show that only FGF1 and FGF2 induce a statistically significant downregulation of these proteins compared to controls. c,d. Increase in the numbers of BrdU + OLs and levels of p21cip1 also show that only FGF1 and FGF2 treatment induces mature OLs to re-enter the cell cycle. Error bars represent SEM; N = 3–7 independent experiments, each performed in triplicate. * p < .05. FGF1, FGF2, FGF4, FGF6, FGF9 were used at 10 ng/ml and FGF5, FGF16, FGF20 at 20 ng/ml. Inset, show representative immunoblots for the expression of MOG, FGFR2, p21cip1 and actin as protein loading control.

Article Snippet: The membranes were blocked for 30 min (Tris buffered saline, 0.2% Tween 20, and 5% non-fat powdered milk or 5% BSA), incubated for 1 hr in primary antibodies: myelin oligodendrocyte glycoprotein (MOG; gift from Dr. C. Linington, Aberdeen, UK); FGFR2, (Santa Cruz Biotech, CA, Cat# sc-6930, RRID:AB_669015); β-actin (Sigma, St Louis, MO); p21cip1 (BD Biosciences, San Jose, CA BD Biosciences Cat# 556431, RRID:AB_396415); mCNP (BioLegend, Lutherville, MD Cat# SMI 91, RRID:AB_2565362).

Techniques: Control, Expressing, BrdU Incorporation Assay, Western Blot

Journal: ASN NEURO

Article Title: Diverse Responses of Oligodendrocytes to Different FGF-Family Members: Uncoupling Structure-Function Relationship Within FGF Subfamilies

doi: 10.1080/17590914.2024.2371163

Figure Lengend Snippet: Effect of different FGFs on mature oligodendrocytes after the addition of exogenous heparin. Mature OLs were grown in the absence (control, open bars) or presence of heparin (black bars) and treated with FGF4, FGF5, FGF6, FGF9, FGF16 or FGF20 for 2 d and analyzed for myelin protein expression and cell cycle re-entry. (a,b) immunoblot analysis and quantification of myelin proteins (MOG, FGFR2) show that FGF4, FGF6, and FGF9 significantly downregulated these proteins in the presence of heparin. c, Quantification of p21cip1 show that in the presence of heparin FGF4, FGF6, and FGF9 cause a significant increase in its levels. Error bars represent SEM; N = 3–4 independent experiments, each performed in triplicate. * p < .05. FGF1, FGF2, FGF4, FGF6, FGF9 were used at 10 ng/ml and FGF5, FGF16, FGF20 at 20 ng/ml. Inset, show representative immunoblots for the expression of MOG, FGFR2, p21cip1 and actin as protein loading control.

Article Snippet: The membranes were blocked for 30 min (Tris buffered saline, 0.2% Tween 20, and 5% non-fat powdered milk or 5% BSA), incubated for 1 hr in primary antibodies: myelin oligodendrocyte glycoprotein (MOG; gift from Dr. C. Linington, Aberdeen, UK); FGFR2, (Santa Cruz Biotech, CA, Cat# sc-6930, RRID:AB_669015); β-actin (Sigma, St Louis, MO); p21cip1 (BD Biosciences, San Jose, CA BD Biosciences Cat# 556431, RRID:AB_396415); mCNP (BioLegend, Lutherville, MD Cat# SMI 91, RRID:AB_2565362).

Techniques: Control, Expressing, Western Blot