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phospho 4ebp1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho 4ebp1
    a-c, Representative images of phosphorylated S6 (p-S6, green), RBPMS (red), and DAPI (blue) staining in whole retinal sections (left), with magnified views (middle), and normalization of p-S6/RBPMS intensity (right) from pre-birth (E18.5) embryos, 5 min, and 12 hr postnatal pups. Scale bars, 200 μm (whole retinal section); 50 μm (magnified views). d, Quantification showing that the percentage of p-S6-positive RGCs significantly decreases after birth. Each circle represents the mean percentage of p-S6-positive RGCs among total RBPMS-positive RGCs in the ganglion cell layer (GCL) of each retina. For each whole retina section, all RGCs were included in the quantification. ****p < 0.0001; N = 4, 3 and 3 biological replicates for pre-birth, 5 min post-birth and 12 hr post-birth groups, respectively; unpaired Welch’s t-test. e, Western blot showing that mTOR activity markers (p-S6, p-S6K, and <t>p-4EBP1)</t> decrease markedly in brain, heart, and hindlimb muscles within 5 min after birth. Quadriceps femoris muscles and biceps femoris muscles from both left and right hindlimbs were pooled together to generate hindlimb muscles sample for each mouse. f, Quantification of Western blot result shown in (e) . *p < 0.05, **p < 0.01, ***p < 0.001; N = 3 and 4 biological replicates for pre-birth and 5 min post birth, respectively; unpaired Welch’s t-test. g, Experimental scheme (left) illustrating conditions with deprivation of maternal nutrient supply or inhibition of breathing onset. Representative images (right) of p-S6, RBPMS, and DAPI staining in whole retinal sections (from newborns at 5 min after birth) and corresponding magnified views under the indicated conditions. 200 μm (whole retinal section); 50 μm (magnified views). h, Quantification showing the percentage of p-S6-positive RGCs under different conditions. At 5 min after birth, deprivation of maternal nutrient supply does not alter p-S6 levels, whereas initiation of breathing significantly decreases mTOR activity. ***p < 0.001; n.s., not significant; N = 5, 3, 3 and 3 biological replicates for (“+ nutrient - breathing”), (“- nutrient - breathing”), (“+ nutrient + breathing”) and (“- nutrient + breathing”) groups, respectively; unpaired Welch’s t-test.
    Phospho 4ebp1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 2478 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/4ebp1/Phospho-4E-BP1+(Thr37%2F46)+Rabbit+mAb/bio_rxiv__64898__2026__10__03__756425-223-11-13
    Average 96 stars, based on 2478 article reviews
    phospho 4ebp1 - by Bioz Stars, 2026-10
    96/100 stars

    Images

    1) Product Images from "The Onset of Breathing at Birth Switches Off Regenerative Capacity"

    Article Title: The Onset of Breathing at Birth Switches Off Regenerative Capacity

    Journal: bioRxiv

    doi: 10.64898/2026.10.03.756425

    a-c, Representative images of phosphorylated S6 (p-S6, green), RBPMS (red), and DAPI (blue) staining in whole retinal sections (left), with magnified views (middle), and normalization of p-S6/RBPMS intensity (right) from pre-birth (E18.5) embryos, 5 min, and 12 hr postnatal pups. Scale bars, 200 μm (whole retinal section); 50 μm (magnified views). d, Quantification showing that the percentage of p-S6-positive RGCs significantly decreases after birth. Each circle represents the mean percentage of p-S6-positive RGCs among total RBPMS-positive RGCs in the ganglion cell layer (GCL) of each retina. For each whole retina section, all RGCs were included in the quantification. ****p < 0.0001; N = 4, 3 and 3 biological replicates for pre-birth, 5 min post-birth and 12 hr post-birth groups, respectively; unpaired Welch’s t-test. e, Western blot showing that mTOR activity markers (p-S6, p-S6K, and p-4EBP1) decrease markedly in brain, heart, and hindlimb muscles within 5 min after birth. Quadriceps femoris muscles and biceps femoris muscles from both left and right hindlimbs were pooled together to generate hindlimb muscles sample for each mouse. f, Quantification of Western blot result shown in (e) . *p < 0.05, **p < 0.01, ***p < 0.001; N = 3 and 4 biological replicates for pre-birth and 5 min post birth, respectively; unpaired Welch’s t-test. g, Experimental scheme (left) illustrating conditions with deprivation of maternal nutrient supply or inhibition of breathing onset. Representative images (right) of p-S6, RBPMS, and DAPI staining in whole retinal sections (from newborns at 5 min after birth) and corresponding magnified views under the indicated conditions. 200 μm (whole retinal section); 50 μm (magnified views). h, Quantification showing the percentage of p-S6-positive RGCs under different conditions. At 5 min after birth, deprivation of maternal nutrient supply does not alter p-S6 levels, whereas initiation of breathing significantly decreases mTOR activity. ***p < 0.001; n.s., not significant; N = 5, 3, 3 and 3 biological replicates for (“+ nutrient - breathing”), (“- nutrient - breathing”), (“+ nutrient + breathing”) and (“- nutrient + breathing”) groups, respectively; unpaired Welch’s t-test.
    Figure Legend Snippet: a-c, Representative images of phosphorylated S6 (p-S6, green), RBPMS (red), and DAPI (blue) staining in whole retinal sections (left), with magnified views (middle), and normalization of p-S6/RBPMS intensity (right) from pre-birth (E18.5) embryos, 5 min, and 12 hr postnatal pups. Scale bars, 200 μm (whole retinal section); 50 μm (magnified views). d, Quantification showing that the percentage of p-S6-positive RGCs significantly decreases after birth. Each circle represents the mean percentage of p-S6-positive RGCs among total RBPMS-positive RGCs in the ganglion cell layer (GCL) of each retina. For each whole retina section, all RGCs were included in the quantification. ****p < 0.0001; N = 4, 3 and 3 biological replicates for pre-birth, 5 min post-birth and 12 hr post-birth groups, respectively; unpaired Welch’s t-test. e, Western blot showing that mTOR activity markers (p-S6, p-S6K, and p-4EBP1) decrease markedly in brain, heart, and hindlimb muscles within 5 min after birth. Quadriceps femoris muscles and biceps femoris muscles from both left and right hindlimbs were pooled together to generate hindlimb muscles sample for each mouse. f, Quantification of Western blot result shown in (e) . *p < 0.05, **p < 0.01, ***p < 0.001; N = 3 and 4 biological replicates for pre-birth and 5 min post birth, respectively; unpaired Welch’s t-test. g, Experimental scheme (left) illustrating conditions with deprivation of maternal nutrient supply or inhibition of breathing onset. Representative images (right) of p-S6, RBPMS, and DAPI staining in whole retinal sections (from newborns at 5 min after birth) and corresponding magnified views under the indicated conditions. 200 μm (whole retinal section); 50 μm (magnified views). h, Quantification showing the percentage of p-S6-positive RGCs under different conditions. At 5 min after birth, deprivation of maternal nutrient supply does not alter p-S6 levels, whereas initiation of breathing significantly decreases mTOR activity. ***p < 0.001; n.s., not significant; N = 5, 3, 3 and 3 biological replicates for (“+ nutrient - breathing”), (“- nutrient - breathing”), (“+ nutrient + breathing”) and (“- nutrient + breathing”) groups, respectively; unpaired Welch’s t-test.

    Techniques Used: Staining, Western Blot, Activity Assay, Muscles, Inhibition

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    Western Blot:

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    a-c, Representative images of phosphorylated S6 (p-S6, green), RBPMS (red), and DAPI (blue) staining in whole retinal sections (left), with magnified views (middle), and normalization of p-S6/RBPMS intensity (right) from pre-birth (E18.5) embryos, 5 min, and 12 hr postnatal pups. Scale bars, 200 μm (whole retinal section); 50 μm (magnified views). d, Quantification showing that the percentage of p-S6-positive RGCs significantly decreases after birth. Each circle represents the mean percentage of p-S6-positive RGCs among total RBPMS-positive RGCs in the ganglion cell layer (GCL) of each retina. For each whole retina section, all RGCs were included in the quantification. ****p < 0.0001; N = 4, 3 and 3 biological replicates for pre-birth, 5 min post-birth and 12 hr post-birth groups, respectively; unpaired Welch’s t-test. e, Western blot showing that mTOR activity markers (p-S6, p-S6K, and <t>p-4EBP1)</t> decrease markedly in brain, heart, and hindlimb muscles within 5 min after birth. Quadriceps femoris muscles and biceps femoris muscles from both left and right hindlimbs were pooled together to generate hindlimb muscles sample for each mouse. f, Quantification of Western blot result shown in (e) . *p < 0.05, **p < 0.01, ***p < 0.001; N = 3 and 4 biological replicates for pre-birth and 5 min post birth, respectively; unpaired Welch’s t-test. g, Experimental scheme (left) illustrating conditions with deprivation of maternal nutrient supply or inhibition of breathing onset. Representative images (right) of p-S6, RBPMS, and DAPI staining in whole retinal sections (from newborns at 5 min after birth) and corresponding magnified views under the indicated conditions. 200 μm (whole retinal section); 50 μm (magnified views). h, Quantification showing the percentage of p-S6-positive RGCs under different conditions. At 5 min after birth, deprivation of maternal nutrient supply does not alter p-S6 levels, whereas initiation of breathing significantly decreases mTOR activity. ***p < 0.001; n.s., not significant; N = 5, 3, 3 and 3 biological replicates for (“+ nutrient - breathing”), (“- nutrient - breathing”), (“+ nutrient + breathing”) and (“- nutrient + breathing”) groups, respectively; unpaired Welch’s t-test.
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    a-c, Representative images of phosphorylated S6 (p-S6, green), RBPMS (red), and DAPI (blue) staining in whole retinal sections (left), with magnified views (middle), and normalization of p-S6/RBPMS intensity (right) from pre-birth (E18.5) embryos, 5 min, and 12 hr postnatal pups. Scale bars, 200 μm (whole retinal section); 50 μm (magnified views). d, Quantification showing that the percentage of p-S6-positive RGCs significantly decreases after birth. Each circle represents the mean percentage of p-S6-positive RGCs among total RBPMS-positive RGCs in the ganglion cell layer (GCL) of each retina. For each whole retina section, all RGCs were included in the quantification. ****p < 0.0001; N = 4, 3 and 3 biological replicates for pre-birth, 5 min post-birth and 12 hr post-birth groups, respectively; unpaired Welch’s t-test. e, Western blot showing that mTOR activity markers (p-S6, p-S6K, and <t>p-4EBP1)</t> decrease markedly in brain, heart, and hindlimb muscles within 5 min after birth. Quadriceps femoris muscles and biceps femoris muscles from both left and right hindlimbs were pooled together to generate hindlimb muscles sample for each mouse. f, Quantification of Western blot result shown in (e) . *p < 0.05, **p < 0.01, ***p < 0.001; N = 3 and 4 biological replicates for pre-birth and 5 min post birth, respectively; unpaired Welch’s t-test. g, Experimental scheme (left) illustrating conditions with deprivation of maternal nutrient supply or inhibition of breathing onset. Representative images (right) of p-S6, RBPMS, and DAPI staining in whole retinal sections (from newborns at 5 min after birth) and corresponding magnified views under the indicated conditions. 200 μm (whole retinal section); 50 μm (magnified views). h, Quantification showing the percentage of p-S6-positive RGCs under different conditions. At 5 min after birth, deprivation of maternal nutrient supply does not alter p-S6 levels, whereas initiation of breathing significantly decreases mTOR activity. ***p < 0.001; n.s., not significant; N = 5, 3, 3 and 3 biological replicates for (“+ nutrient - breathing”), (“- nutrient - breathing”), (“+ nutrient + breathing”) and (“- nutrient + breathing”) groups, respectively; unpaired Welch’s t-test.
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    a-c, Representative images of phosphorylated S6 (p-S6, green), RBPMS (red), and DAPI (blue) staining in whole retinal sections (left), with magnified views (middle), and normalization of p-S6/RBPMS intensity (right) from pre-birth (E18.5) embryos, 5 min, and 12 hr postnatal pups. Scale bars, 200 μm (whole retinal section); 50 μm (magnified views). d, Quantification showing that the percentage of p-S6-positive RGCs significantly decreases after birth. Each circle represents the mean percentage of p-S6-positive RGCs among total RBPMS-positive RGCs in the ganglion cell layer (GCL) of each retina. For each whole retina section, all RGCs were included in the quantification. ****p < 0.0001; N = 4, 3 and 3 biological replicates for pre-birth, 5 min post-birth and 12 hr post-birth groups, respectively; unpaired Welch’s t-test. e, Western blot showing that mTOR activity markers (p-S6, p-S6K, and <t>p-4EBP1)</t> decrease markedly in brain, heart, and hindlimb muscles within 5 min after birth. Quadriceps femoris muscles and biceps femoris muscles from both left and right hindlimbs were pooled together to generate hindlimb muscles sample for each mouse. f, Quantification of Western blot result shown in (e) . *p < 0.05, **p < 0.01, ***p < 0.001; N = 3 and 4 biological replicates for pre-birth and 5 min post birth, respectively; unpaired Welch’s t-test. g, Experimental scheme (left) illustrating conditions with deprivation of maternal nutrient supply or inhibition of breathing onset. Representative images (right) of p-S6, RBPMS, and DAPI staining in whole retinal sections (from newborns at 5 min after birth) and corresponding magnified views under the indicated conditions. 200 μm (whole retinal section); 50 μm (magnified views). h, Quantification showing the percentage of p-S6-positive RGCs under different conditions. At 5 min after birth, deprivation of maternal nutrient supply does not alter p-S6 levels, whereas initiation of breathing significantly decreases mTOR activity. ***p < 0.001; n.s., not significant; N = 5, 3, 3 and 3 biological replicates for (“+ nutrient - breathing”), (“- nutrient - breathing”), (“+ nutrient + breathing”) and (“- nutrient + breathing”) groups, respectively; unpaired Welch’s t-test.
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    a-c, Representative images of phosphorylated S6 (p-S6, green), RBPMS (red), and DAPI (blue) staining in whole retinal sections (left), with magnified views (middle), and normalization of p-S6/RBPMS intensity (right) from pre-birth (E18.5) embryos, 5 min, and 12 hr postnatal pups. Scale bars, 200 μm (whole retinal section); 50 μm (magnified views). d, Quantification showing that the percentage of p-S6-positive RGCs significantly decreases after birth. Each circle represents the mean percentage of p-S6-positive RGCs among total RBPMS-positive RGCs in the ganglion cell layer (GCL) of each retina. For each whole retina section, all RGCs were included in the quantification. ****p < 0.0001; N = 4, 3 and 3 biological replicates for pre-birth, 5 min post-birth and 12 hr post-birth groups, respectively; unpaired Welch’s t-test. e, Western blot showing that mTOR activity markers (p-S6, p-S6K, and <t>p-4EBP1)</t> decrease markedly in brain, heart, and hindlimb muscles within 5 min after birth. Quadriceps femoris muscles and biceps femoris muscles from both left and right hindlimbs were pooled together to generate hindlimb muscles sample for each mouse. f, Quantification of Western blot result shown in (e) . *p < 0.05, **p < 0.01, ***p < 0.001; N = 3 and 4 biological replicates for pre-birth and 5 min post birth, respectively; unpaired Welch’s t-test. g, Experimental scheme (left) illustrating conditions with deprivation of maternal nutrient supply or inhibition of breathing onset. Representative images (right) of p-S6, RBPMS, and DAPI staining in whole retinal sections (from newborns at 5 min after birth) and corresponding magnified views under the indicated conditions. 200 μm (whole retinal section); 50 μm (magnified views). h, Quantification showing the percentage of p-S6-positive RGCs under different conditions. At 5 min after birth, deprivation of maternal nutrient supply does not alter p-S6 levels, whereas initiation of breathing significantly decreases mTOR activity. ***p < 0.001; n.s., not significant; N = 5, 3, 3 and 3 biological replicates for (“+ nutrient - breathing”), (“- nutrient - breathing”), (“+ nutrient + breathing”) and (“- nutrient + breathing”) groups, respectively; unpaired Welch’s t-test.
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    a-c, Representative images of phosphorylated S6 (p-S6, green), RBPMS (red), and DAPI (blue) staining in whole retinal sections (left), with magnified views (middle), and normalization of p-S6/RBPMS intensity (right) from pre-birth (E18.5) embryos, 5 min, and 12 hr postnatal pups. Scale bars, 200 μm (whole retinal section); 50 μm (magnified views). d, Quantification showing that the percentage of p-S6-positive RGCs significantly decreases after birth. Each circle represents the mean percentage of p-S6-positive RGCs among total RBPMS-positive RGCs in the ganglion cell layer (GCL) of each retina. For each whole retina section, all RGCs were included in the quantification. ****p < 0.0001; N = 4, 3 and 3 biological replicates for pre-birth, 5 min post-birth and 12 hr post-birth groups, respectively; unpaired Welch’s t-test. e, Western blot showing that mTOR activity markers (p-S6, p-S6K, and <t>p-4EBP1)</t> decrease markedly in brain, heart, and hindlimb muscles within 5 min after birth. Quadriceps femoris muscles and biceps femoris muscles from both left and right hindlimbs were pooled together to generate hindlimb muscles sample for each mouse. f, Quantification of Western blot result shown in (e) . *p < 0.05, **p < 0.01, ***p < 0.001; N = 3 and 4 biological replicates for pre-birth and 5 min post birth, respectively; unpaired Welch’s t-test. g, Experimental scheme (left) illustrating conditions with deprivation of maternal nutrient supply or inhibition of breathing onset. Representative images (right) of p-S6, RBPMS, and DAPI staining in whole retinal sections (from newborns at 5 min after birth) and corresponding magnified views under the indicated conditions. 200 μm (whole retinal section); 50 μm (magnified views). h, Quantification showing the percentage of p-S6-positive RGCs under different conditions. At 5 min after birth, deprivation of maternal nutrient supply does not alter p-S6 levels, whereas initiation of breathing significantly decreases mTOR activity. ***p < 0.001; n.s., not significant; N = 5, 3, 3 and 3 biological replicates for (“+ nutrient - breathing”), (“- nutrient - breathing”), (“+ nutrient + breathing”) and (“- nutrient + breathing”) groups, respectively; unpaired Welch’s t-test.
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    a-c, Representative images of phosphorylated S6 (p-S6, green), RBPMS (red), and DAPI (blue) staining in whole retinal sections (left), with magnified views (middle), and normalization of p-S6/RBPMS intensity (right) from pre-birth (E18.5) embryos, 5 min, and 12 hr postnatal pups. Scale bars, 200 μm (whole retinal section); 50 μm (magnified views). d, Quantification showing that the percentage of p-S6-positive RGCs significantly decreases after birth. Each circle represents the mean percentage of p-S6-positive RGCs among total RBPMS-positive RGCs in the ganglion cell layer (GCL) of each retina. For each whole retina section, all RGCs were included in the quantification. ****p < 0.0001; N = 4, 3 and 3 biological replicates for pre-birth, 5 min post-birth and 12 hr post-birth groups, respectively; unpaired Welch’s t-test. e, Western blot showing that mTOR activity markers (p-S6, p-S6K, and <t>p-4EBP1)</t> decrease markedly in brain, heart, and hindlimb muscles within 5 min after birth. Quadriceps femoris muscles and biceps femoris muscles from both left and right hindlimbs were pooled together to generate hindlimb muscles sample for each mouse. f, Quantification of Western blot result shown in (e) . *p < 0.05, **p < 0.01, ***p < 0.001; N = 3 and 4 biological replicates for pre-birth and 5 min post birth, respectively; unpaired Welch’s t-test. g, Experimental scheme (left) illustrating conditions with deprivation of maternal nutrient supply or inhibition of breathing onset. Representative images (right) of p-S6, RBPMS, and DAPI staining in whole retinal sections (from newborns at 5 min after birth) and corresponding magnified views under the indicated conditions. 200 μm (whole retinal section); 50 μm (magnified views). h, Quantification showing the percentage of p-S6-positive RGCs under different conditions. At 5 min after birth, deprivation of maternal nutrient supply does not alter p-S6 levels, whereas initiation of breathing significantly decreases mTOR activity. ***p < 0.001; n.s., not significant; N = 5, 3, 3 and 3 biological replicates for (“+ nutrient - breathing”), (“- nutrient - breathing”), (“+ nutrient + breathing”) and (“- nutrient + breathing”) groups, respectively; unpaired Welch’s t-test.
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    a-c, Representative images of phosphorylated S6 (p-S6, green), RBPMS (red), and DAPI (blue) staining in whole retinal sections (left), with magnified views (middle), and normalization of p-S6/RBPMS intensity (right) from pre-birth (E18.5) embryos, 5 min, and 12 hr postnatal pups. Scale bars, 200 μm (whole retinal section); 50 μm (magnified views). d, Quantification showing that the percentage of p-S6-positive RGCs significantly decreases after birth. Each circle represents the mean percentage of p-S6-positive RGCs among total RBPMS-positive RGCs in the ganglion cell layer (GCL) of each retina. For each whole retina section, all RGCs were included in the quantification. ****p < 0.0001; N = 4, 3 and 3 biological replicates for pre-birth, 5 min post-birth and 12 hr post-birth groups, respectively; unpaired Welch’s t-test. e, Western blot showing that mTOR activity markers (p-S6, p-S6K, and <t>p-4EBP1)</t> decrease markedly in brain, heart, and hindlimb muscles within 5 min after birth. Quadriceps femoris muscles and biceps femoris muscles from both left and right hindlimbs were pooled together to generate hindlimb muscles sample for each mouse. f, Quantification of Western blot result shown in (e) . *p < 0.05, **p < 0.01, ***p < 0.001; N = 3 and 4 biological replicates for pre-birth and 5 min post birth, respectively; unpaired Welch’s t-test. g, Experimental scheme (left) illustrating conditions with deprivation of maternal nutrient supply or inhibition of breathing onset. Representative images (right) of p-S6, RBPMS, and DAPI staining in whole retinal sections (from newborns at 5 min after birth) and corresponding magnified views under the indicated conditions. 200 μm (whole retinal section); 50 μm (magnified views). h, Quantification showing the percentage of p-S6-positive RGCs under different conditions. At 5 min after birth, deprivation of maternal nutrient supply does not alter p-S6 levels, whereas initiation of breathing significantly decreases mTOR activity. ***p < 0.001; n.s., not significant; N = 5, 3, 3 and 3 biological replicates for (“+ nutrient - breathing”), (“- nutrient - breathing”), (“+ nutrient + breathing”) and (“- nutrient + breathing”) groups, respectively; unpaired Welch’s t-test.
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    Image Search Results


    a-c, Representative images of phosphorylated S6 (p-S6, green), RBPMS (red), and DAPI (blue) staining in whole retinal sections (left), with magnified views (middle), and normalization of p-S6/RBPMS intensity (right) from pre-birth (E18.5) embryos, 5 min, and 12 hr postnatal pups. Scale bars, 200 μm (whole retinal section); 50 μm (magnified views). d, Quantification showing that the percentage of p-S6-positive RGCs significantly decreases after birth. Each circle represents the mean percentage of p-S6-positive RGCs among total RBPMS-positive RGCs in the ganglion cell layer (GCL) of each retina. For each whole retina section, all RGCs were included in the quantification. ****p < 0.0001; N = 4, 3 and 3 biological replicates for pre-birth, 5 min post-birth and 12 hr post-birth groups, respectively; unpaired Welch’s t-test. e, Western blot showing that mTOR activity markers (p-S6, p-S6K, and p-4EBP1) decrease markedly in brain, heart, and hindlimb muscles within 5 min after birth. Quadriceps femoris muscles and biceps femoris muscles from both left and right hindlimbs were pooled together to generate hindlimb muscles sample for each mouse. f, Quantification of Western blot result shown in (e) . *p < 0.05, **p < 0.01, ***p < 0.001; N = 3 and 4 biological replicates for pre-birth and 5 min post birth, respectively; unpaired Welch’s t-test. g, Experimental scheme (left) illustrating conditions with deprivation of maternal nutrient supply or inhibition of breathing onset. Representative images (right) of p-S6, RBPMS, and DAPI staining in whole retinal sections (from newborns at 5 min after birth) and corresponding magnified views under the indicated conditions. 200 μm (whole retinal section); 50 μm (magnified views). h, Quantification showing the percentage of p-S6-positive RGCs under different conditions. At 5 min after birth, deprivation of maternal nutrient supply does not alter p-S6 levels, whereas initiation of breathing significantly decreases mTOR activity. ***p < 0.001; n.s., not significant; N = 5, 3, 3 and 3 biological replicates for (“+ nutrient - breathing”), (“- nutrient - breathing”), (“+ nutrient + breathing”) and (“- nutrient + breathing”) groups, respectively; unpaired Welch’s t-test.

    Journal: bioRxiv

    Article Title: The Onset of Breathing at Birth Switches Off Regenerative Capacity

    doi: 10.64898/2026.10.03.756425

    Figure Lengend Snippet: a-c, Representative images of phosphorylated S6 (p-S6, green), RBPMS (red), and DAPI (blue) staining in whole retinal sections (left), with magnified views (middle), and normalization of p-S6/RBPMS intensity (right) from pre-birth (E18.5) embryos, 5 min, and 12 hr postnatal pups. Scale bars, 200 μm (whole retinal section); 50 μm (magnified views). d, Quantification showing that the percentage of p-S6-positive RGCs significantly decreases after birth. Each circle represents the mean percentage of p-S6-positive RGCs among total RBPMS-positive RGCs in the ganglion cell layer (GCL) of each retina. For each whole retina section, all RGCs were included in the quantification. ****p < 0.0001; N = 4, 3 and 3 biological replicates for pre-birth, 5 min post-birth and 12 hr post-birth groups, respectively; unpaired Welch’s t-test. e, Western blot showing that mTOR activity markers (p-S6, p-S6K, and p-4EBP1) decrease markedly in brain, heart, and hindlimb muscles within 5 min after birth. Quadriceps femoris muscles and biceps femoris muscles from both left and right hindlimbs were pooled together to generate hindlimb muscles sample for each mouse. f, Quantification of Western blot result shown in (e) . *p < 0.05, **p < 0.01, ***p < 0.001; N = 3 and 4 biological replicates for pre-birth and 5 min post birth, respectively; unpaired Welch’s t-test. g, Experimental scheme (left) illustrating conditions with deprivation of maternal nutrient supply or inhibition of breathing onset. Representative images (right) of p-S6, RBPMS, and DAPI staining in whole retinal sections (from newborns at 5 min after birth) and corresponding magnified views under the indicated conditions. 200 μm (whole retinal section); 50 μm (magnified views). h, Quantification showing the percentage of p-S6-positive RGCs under different conditions. At 5 min after birth, deprivation of maternal nutrient supply does not alter p-S6 levels, whereas initiation of breathing significantly decreases mTOR activity. ***p < 0.001; n.s., not significant; N = 5, 3, 3 and 3 biological replicates for (“+ nutrient - breathing”), (“- nutrient - breathing”), (“+ nutrient + breathing”) and (“- nutrient + breathing”) groups, respectively; unpaired Welch’s t-test.

    Article Snippet: Primary antibodies against phospho-S6 (Ser235/236, CST 4858S), phospho-S6K (Thr389, CST 9234T), phospho-4EBP1 (Thr37/46, CST 2855T), total S6 (CST 2217T), S6K (CST 9202S), 4EBP1(CST 9644T) and actin (CST 4970T) were obtained from Cell Signaling Technology.

    Techniques: Staining, Western Blot, Activity Assay, Muscles, Inhibition