Extended Data Fig. 3: APJ signaling specifically stimulates oligodendrocyte differentiation. | Nature Aging

Extended Data Fig. 3: APJ signaling specifically stimulates oligodendrocyte differentiation.

From: Age-dependent decline in remyelination capacity is mediated by apelin–APJ signaling

Extended Data Fig. 3

a, Representative images of MBP expression in mouse oligodendrocyte culture. Cells were transfected with Cy5-labeled indicated siRNA and then cultured in the presence of ML233 for an additional 3 days. Quantitation of the MBP-positive area under the culture conditions indicated in the images (n = 3). Each data point represents biologically independent experiments. P = 0.007, 0.009 (left to right). Closed arrowheads, MBP-expressing cells transfected with siRNA; open arrowheads, MBP-negative cells transfected with siRNA. b, Immunoblot of Myrf expression in the nuclear and total fractions. Samples were prepared from 3-week-old mice and aged mice (n = 3), P = 0.01. A cropped image of the gel is presented (see Source Date 2 for full image). c, Immunoblot of Myrf expression in the nuclear and total fractions. Samples were prepared from wild-type mice and APJ KO mice (n = 3), P = 0.014. A cropped image of the gel is presented (see Source Date 2 for full image). d, Representative images of MBP expression in mouse oligodendrocyte culture. Cells were treated with the PKC inhibitor Gö6983 (10 μM) for 30 min and then cultured in the presence of ML233 for an additional 3 days. Quantitation of the MBP-positive area under the culture conditions indicated in the images (n = 4). Each data point represents biologically independent experiments. P = 0.004, 0.002 (left to right). e, Representative images of MBP expression in mouse oligodendrocyte culture. Cells were treated with the PI3K inhibitor LY294002 (10 μM) for 30 min and then cultured in the presence of ML233 for an additional 3 days. Quantitation of the MBP-positive area under the culture conditions indicated in the images (n = 4). Each data point represents biologically independent experiments. P < 0.001, 0.002 (left to right). f, Representative images of MBP expression in mouse oligodendrocyte culture. Cells were treated with the MEK inhibitor U0126 (10 μM) for 30 min and then cultured in the presence of ML233 for an additional 3 days. Quantitation of the MBP-positive area under the culture conditions indicated in the images (n = 4). Each data point represents biologically independent experiments. P = 0.004, 0.006 (left to right). g, Representative images of MBP expression in mouse oligodendrocyte culture. Cells were cultured in the neuronal supernatant for 3 days. Neuronal supernatant was collected from the cortical neuron culture with ML233 pretreatment. Quantitation of the MBP-positive area under the culture conditions indicated in the images (n = 4). Each data point represents biologically independent experiments. P = 0.31. Scale bars, 100 μm. Data are the mean ± s.e.m. and were assessed using one‐way ANOVA with post hoc Tukey’s multiple comparison test (a, d, e, and f) and two-tailed Welch’s t-test (b, c, and g).

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