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rheb1  (R&D Systems)


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    Structured Review

    R&D Systems rheb1
    Rheb1, supplied by R&D Systems, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rheb1/Human%2FMouse%2FRat+Rheb+Antibody/10__1523_slash_JNEUROSCI__0449___21__2022_ascii32_-78-24-28
    Average 91 stars, based on 3 article reviews
    rheb1 - by Bioz Stars, 2026-10
    91/100 stars

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    Related Articles

    Knock-Out:

    Article Title: A Farnesyltransferase Inhibitor Restores Cognitive Deficits in<i>Tsc2</i><sup>+/–</sup>Mice through Inhibition of Rheb1
    Article Snippet: HEK293 T cells, cultured hippocampal neurons, and mouse brains and livers were homogenized [40 mM Tris-HCl, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% Triton X-100, protease inhibitor cocktail tablet (Complete EDTA-free; catalog #11873580001, Roche) and 1 mM phenylmethanelsulfonyl fluoride (catalog #P7626, Sigma-Aldrich)] and centrifuged at 17,900 g at 4°C.and centrifuged at 17,900 g at 4°C. ... The resultant soluble and insoluble fractions were immunoblotted as described previously (Sugiura et al., 2015), and primary antibodies against FLAG (1:800; catalog #F3165, Sigma-Aldrich), Rheb1 [1:1000; catalog #MAB3426, R&D Systems (recognizing whole Rheb); 1:1000; catalog #13879; Cell Signaling Technology (preferably recognizing unfarnesylated Rheb)], a/b -tubulin (1:1000; catalog #2148, Cell Signaling Technology), phospho (p)-ERK1/2 (Thr202/Tyr204; 1:1000; catalog #9101, Cell Signaling Technology), ERK1/2 (1:1000; catalog #9102, Cell Signaling Technology), syntenin1 (1:400; catalog #sc-100336, Santa Cruz Biotechnology), p-S6 (Ser240/244; 1:1000; catalog #5364, Cell Signaling Technology), and S6 (1:1000; catalog #2317, Cell Signaling Technology), and HRP-conjugated secondary antibodies (1:6000; catalog #170–6515, BIO-RAD) were used.. Quantitative analysis of dendritic arc immunofluorescence.Quantitative analysis of dendritic arc immunofluorescence.

    Expressing:

    Article Title: A Farnesyltransferase Inhibitor Restores Cognitive Deficits in<i>Tsc2</i><sup>+/–</sup>Mice through Inhibition of Rheb1
    Article Snippet: HEK293 T cells, cultured hippocampal neurons, and mouse brains and livers were homogenized [40 mM Tris-HCl, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% Triton X-100, protease inhibitor cocktail tablet (Complete EDTA-free; catalog #11873580001, Roche) and 1 mM phenylmethanelsulfonyl fluoride (catalog #P7626, Sigma-Aldrich)] and centrifuged at 17,900 g at 4°C.and centrifuged at 17,900 g at 4°C. ... The resultant soluble and insoluble fractions were immunoblotted as described previously (Sugiura et al., 2015), and primary antibodies against FLAG (1:800; catalog #F3165, Sigma-Aldrich), Rheb1 [1:1000; catalog #MAB3426, R&D Systems (recognizing whole Rheb); 1:1000; catalog #13879; Cell Signaling Technology (preferably recognizing unfarnesylated Rheb)], a/b -tubulin (1:1000; catalog #2148, Cell Signaling Technology), phospho (p)-ERK1/2 (Thr202/Tyr204; 1:1000; catalog #9101, Cell Signaling Technology), ERK1/2 (1:1000; catalog #9102, Cell Signaling Technology), syntenin1 (1:400; catalog #sc-100336, Santa Cruz Biotechnology), p-S6 (Ser240/244; 1:1000; catalog #5364, Cell Signaling Technology), and S6 (1:1000; catalog #2317, Cell Signaling Technology), and HRP-conjugated secondary antibodies (1:6000; catalog #170–6515, BIO-RAD) were used.. Quantitative analysis of dendritic arc immunofluorescence.Quantitative analysis of dendritic arc immunofluorescence.

    Cell Culture:

    Article Title: A Farnesyltransferase Inhibitor Restores Cognitive Deficits in<i>Tsc2</i><sup>+/–</sup>Mice through Inhibition of Rheb1
    Article Snippet: HEK293 T cells, cultured hippocampal neurons, and mouse brains and livers were homogenized [40 mM Tris-HCl, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% Triton X-100, protease inhibitor cocktail tablet (Complete EDTA-free; catalog #11873580001, Roche) and 1 mM phenylmethanelsulfonyl fluoride (catalog #P7626, Sigma-Aldrich)] and centrifuged at 17,900 g at 4°C.and centrifuged at 17,900 g at 4°C. ... The resultant soluble and insoluble fractions were immunoblotted as described previously (Sugiura et al., 2015), and primary antibodies against FLAG (1:800; catalog #F3165, Sigma-Aldrich), Rheb1 [1:1000; catalog #MAB3426, R&D Systems (recognizing whole Rheb); 1:1000; catalog #13879; Cell Signaling Technology (preferably recognizing unfarnesylated Rheb)], a/b -tubulin (1:1000; catalog #2148, Cell Signaling Technology), phospho (p)-ERK1/2 (Thr202/Tyr204; 1:1000; catalog #9101, Cell Signaling Technology), ERK1/2 (1:1000; catalog #9102, Cell Signaling Technology), syntenin1 (1:400; catalog #sc-100336, Santa Cruz Biotechnology), p-S6 (Ser240/244; 1:1000; catalog #5364, Cell Signaling Technology), and S6 (1:1000; catalog #2317, Cell Signaling Technology), and HRP-conjugated secondary antibodies (1:6000; catalog #170–6515, BIO-RAD) were used.. Quantitative analysis of dendritic arc immunofluorescence.Quantitative analysis of dendritic arc immunofluorescence.

    Immunodetection:

    Article Title: A Farnesyltransferase Inhibitor Restores Cognitive Deficits in<i>Tsc2</i><sup>+/–</sup>Mice through Inhibition of Rheb1
    Article Snippet: HEK293 T cells, cultured hippocampal neurons, and mouse brains and livers were homogenized [40 mM Tris-HCl, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% Triton X-100, protease inhibitor cocktail tablet (Complete EDTA-free; catalog #11873580001, Roche) and 1 mM phenylmethanelsulfonyl fluoride (catalog #P7626, Sigma-Aldrich)] and centrifuged at 17,900 g at 4°C.and centrifuged at 17,900 g at 4°C. ... The resultant soluble and insoluble fractions were immunoblotted as described previously (Sugiura et al., 2015), and primary antibodies against FLAG (1:800; catalog #F3165, Sigma-Aldrich), Rheb1 [1:1000; catalog #MAB3426, R&D Systems (recognizing whole Rheb); 1:1000; catalog #13879; Cell Signaling Technology (preferably recognizing unfarnesylated Rheb)], a/b -tubulin (1:1000; catalog #2148, Cell Signaling Technology), phospho (p)-ERK1/2 (Thr202/Tyr204; 1:1000; catalog #9101, Cell Signaling Technology), ERK1/2 (1:1000; catalog #9102, Cell Signaling Technology), syntenin1 (1:400; catalog #sc-100336, Santa Cruz Biotechnology), p-S6 (Ser240/244; 1:1000; catalog #5364, Cell Signaling Technology), and S6 (1:1000; catalog #2317, Cell Signaling Technology), and HRP-conjugated secondary antibodies (1:6000; catalog #170–6515, BIO-RAD) were used.. Quantitative analysis of dendritic arc immunofluorescence.Quantitative analysis of dendritic arc immunofluorescence.

    Modification:

    Article Title: A Farnesyltransferase Inhibitor Restores Cognitive Deficits in<i>Tsc2</i><sup>+/–</sup>Mice through Inhibition of Rheb1
    Article Snippet: HEK293 T cells, cultured hippocampal neurons, and mouse brains and livers were homogenized [40 mM Tris-HCl, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% Triton X-100, protease inhibitor cocktail tablet (Complete EDTA-free; catalog #11873580001, Roche) and 1 mM phenylmethanelsulfonyl fluoride (catalog #P7626, Sigma-Aldrich)] and centrifuged at 17,900 g at 4°C.and centrifuged at 17,900 g at 4°C. ... The resultant soluble and insoluble fractions were immunoblotted as described previously (Sugiura et al., 2015), and primary antibodies against FLAG (1:800; catalog #F3165, Sigma-Aldrich), Rheb1 [1:1000; catalog #MAB3426, R&D Systems (recognizing whole Rheb); 1:1000; catalog #13879; Cell Signaling Technology (preferably recognizing unfarnesylated Rheb)], a/b -tubulin (1:1000; catalog #2148, Cell Signaling Technology), phospho (p)-ERK1/2 (Thr202/Tyr204; 1:1000; catalog #9101, Cell Signaling Technology), ERK1/2 (1:1000; catalog #9102, Cell Signaling Technology), syntenin1 (1:400; catalog #sc-100336, Santa Cruz Biotechnology), p-S6 (Ser240/244; 1:1000; catalog #5364, Cell Signaling Technology), and S6 (1:1000; catalog #2317, Cell Signaling Technology), and HRP-conjugated secondary antibodies (1:6000; catalog #170–6515, BIO-RAD) were used.. Quantitative analysis of dendritic arc immunofluorescence.Quantitative analysis of dendritic arc immunofluorescence.

    SDS Page:

    Article Title: A Farnesyltransferase Inhibitor Restores Cognitive Deficits in<i>Tsc2</i><sup>+/–</sup>Mice through Inhibition of Rheb1
    Article Snippet: HEK293 T cells, cultured hippocampal neurons, and mouse brains and livers were homogenized [40 mM Tris-HCl, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% Triton X-100, protease inhibitor cocktail tablet (Complete EDTA-free; catalog #11873580001, Roche) and 1 mM phenylmethanelsulfonyl fluoride (catalog #P7626, Sigma-Aldrich)] and centrifuged at 17,900 g at 4°C.and centrifuged at 17,900 g at 4°C. ... The resultant soluble and insoluble fractions were immunoblotted as described previously (Sugiura et al., 2015), and primary antibodies against FLAG (1:800; catalog #F3165, Sigma-Aldrich), Rheb1 [1:1000; catalog #MAB3426, R&D Systems (recognizing whole Rheb); 1:1000; catalog #13879; Cell Signaling Technology (preferably recognizing unfarnesylated Rheb)], a/b -tubulin (1:1000; catalog #2148, Cell Signaling Technology), phospho (p)-ERK1/2 (Thr202/Tyr204; 1:1000; catalog #9101, Cell Signaling Technology), ERK1/2 (1:1000; catalog #9102, Cell Signaling Technology), syntenin1 (1:400; catalog #sc-100336, Santa Cruz Biotechnology), p-S6 (Ser240/244; 1:1000; catalog #5364, Cell Signaling Technology), and S6 (1:1000; catalog #2317, Cell Signaling Technology), and HRP-conjugated secondary antibodies (1:6000; catalog #170–6515, BIO-RAD) were used.. Quantitative analysis of dendritic arc immunofluorescence.Quantitative analysis of dendritic arc immunofluorescence.

    Transfection:

    Article Title: A Farnesyltransferase Inhibitor Restores Cognitive Deficits in<i>Tsc2</i><sup>+/–</sup>Mice through Inhibition of Rheb1
    Article Snippet: HEK293 T cells, cultured hippocampal neurons, and mouse brains and livers were homogenized [40 mM Tris-HCl, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% Triton X-100, protease inhibitor cocktail tablet (Complete EDTA-free; catalog #11873580001, Roche) and 1 mM phenylmethanelsulfonyl fluoride (catalog #P7626, Sigma-Aldrich)] and centrifuged at 17,900 g at 4°C.and centrifuged at 17,900 g at 4°C. ... The resultant soluble and insoluble fractions were immunoblotted as described previously (Sugiura et al., 2015), and primary antibodies against FLAG (1:800; catalog #F3165, Sigma-Aldrich), Rheb1 [1:1000; catalog #MAB3426, R&D Systems (recognizing whole Rheb); 1:1000; catalog #13879; Cell Signaling Technology (preferably recognizing unfarnesylated Rheb)], a/b -tubulin (1:1000; catalog #2148, Cell Signaling Technology), phospho (p)-ERK1/2 (Thr202/Tyr204; 1:1000; catalog #9101, Cell Signaling Technology), ERK1/2 (1:1000; catalog #9102, Cell Signaling Technology), syntenin1 (1:400; catalog #sc-100336, Santa Cruz Biotechnology), p-S6 (Ser240/244; 1:1000; catalog #5364, Cell Signaling Technology), and S6 (1:1000; catalog #2317, Cell Signaling Technology), and HRP-conjugated secondary antibodies (1:6000; catalog #170–6515, BIO-RAD) were used.. Quantitative analysis of dendritic arc immunofluorescence.Quantitative analysis of dendritic arc immunofluorescence.

    Incubation:

    Article Title: A Farnesyltransferase Inhibitor Restores Cognitive Deficits in<i>Tsc2</i><sup>+/–</sup>Mice through Inhibition of Rheb1
    Article Snippet: HEK293 T cells, cultured hippocampal neurons, and mouse brains and livers were homogenized [40 mM Tris-HCl, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% Triton X-100, protease inhibitor cocktail tablet (Complete EDTA-free; catalog #11873580001, Roche) and 1 mM phenylmethanelsulfonyl fluoride (catalog #P7626, Sigma-Aldrich)] and centrifuged at 17,900 g at 4°C.and centrifuged at 17,900 g at 4°C. ... The resultant soluble and insoluble fractions were immunoblotted as described previously (Sugiura et al., 2015), and primary antibodies against FLAG (1:800; catalog #F3165, Sigma-Aldrich), Rheb1 [1:1000; catalog #MAB3426, R&D Systems (recognizing whole Rheb); 1:1000; catalog #13879; Cell Signaling Technology (preferably recognizing unfarnesylated Rheb)], a/b -tubulin (1:1000; catalog #2148, Cell Signaling Technology), phospho (p)-ERK1/2 (Thr202/Tyr204; 1:1000; catalog #9101, Cell Signaling Technology), ERK1/2 (1:1000; catalog #9102, Cell Signaling Technology), syntenin1 (1:400; catalog #sc-100336, Santa Cruz Biotechnology), p-S6 (Ser240/244; 1:1000; catalog #5364, Cell Signaling Technology), and S6 (1:1000; catalog #2317, Cell Signaling Technology), and HRP-conjugated secondary antibodies (1:6000; catalog #170–6515, BIO-RAD) were used.. Quantitative analysis of dendritic arc immunofluorescence.Quantitative analysis of dendritic arc immunofluorescence.

    Two Tailed Test:

    Article Title: A Farnesyltransferase Inhibitor Restores Cognitive Deficits in<i>Tsc2</i><sup>+/–</sup>Mice through Inhibition of Rheb1
    Article Snippet: HEK293 T cells, cultured hippocampal neurons, and mouse brains and livers were homogenized [40 mM Tris-HCl, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% Triton X-100, protease inhibitor cocktail tablet (Complete EDTA-free; catalog #11873580001, Roche) and 1 mM phenylmethanelsulfonyl fluoride (catalog #P7626, Sigma-Aldrich)] and centrifuged at 17,900 g at 4°C.and centrifuged at 17,900 g at 4°C. ... The resultant soluble and insoluble fractions were immunoblotted as described previously (Sugiura et al., 2015), and primary antibodies against FLAG (1:800; catalog #F3165, Sigma-Aldrich), Rheb1 [1:1000; catalog #MAB3426, R&D Systems (recognizing whole Rheb); 1:1000; catalog #13879; Cell Signaling Technology (preferably recognizing unfarnesylated Rheb)], a/b -tubulin (1:1000; catalog #2148, Cell Signaling Technology), phospho (p)-ERK1/2 (Thr202/Tyr204; 1:1000; catalog #9101, Cell Signaling Technology), ERK1/2 (1:1000; catalog #9102, Cell Signaling Technology), syntenin1 (1:400; catalog #sc-100336, Santa Cruz Biotechnology), p-S6 (Ser240/244; 1:1000; catalog #5364, Cell Signaling Technology), and S6 (1:1000; catalog #2317, Cell Signaling Technology), and HRP-conjugated secondary antibodies (1:6000; catalog #170–6515, BIO-RAD) were used.. Quantitative analysis of dendritic arc immunofluorescence.Quantitative analysis of dendritic arc immunofluorescence.

    Solubility:

    Article Title: A Farnesyltransferase Inhibitor Restores Cognitive Deficits in<i>Tsc2</i><sup>+/–</sup>Mice through Inhibition of Rheb1
    Article Snippet: HEK293 T cells, cultured hippocampal neurons, and mouse brains and livers were homogenized [40 mM Tris-HCl, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% Triton X-100, protease inhibitor cocktail tablet (Complete EDTA-free; catalog #11873580001, Roche) and 1 mM phenylmethanelsulfonyl fluoride (catalog #P7626, Sigma-Aldrich)] and centrifuged at 17,900 g at 4°C.and centrifuged at 17,900 g at 4°C. ... The resultant soluble and insoluble fractions were immunoblotted as described previously (Sugiura et al., 2015), and primary antibodies against FLAG (1:800; catalog #F3165, Sigma-Aldrich), Rheb1 [1:1000; catalog #MAB3426, R&D Systems (recognizing whole Rheb); 1:1000; catalog #13879; Cell Signaling Technology (preferably recognizing unfarnesylated Rheb)], a/b -tubulin (1:1000; catalog #2148, Cell Signaling Technology), phospho (p)-ERK1/2 (Thr202/Tyr204; 1:1000; catalog #9101, Cell Signaling Technology), ERK1/2 (1:1000; catalog #9102, Cell Signaling Technology), syntenin1 (1:400; catalog #sc-100336, Santa Cruz Biotechnology), p-S6 (Ser240/244; 1:1000; catalog #5364, Cell Signaling Technology), and S6 (1:1000; catalog #2317, Cell Signaling Technology), and HRP-conjugated secondary antibodies (1:6000; catalog #170–6515, BIO-RAD) were used.. Quantitative analysis of dendritic arc immunofluorescence.Quantitative analysis of dendritic arc immunofluorescence.

    Activation Assay:

    Article Title: A Farnesyltransferase Inhibitor Restores Cognitive Deficits in<i>Tsc2</i><sup>+/–</sup>Mice through Inhibition of Rheb1
    Article Snippet: HEK293 T cells, cultured hippocampal neurons, and mouse brains and livers were homogenized [40 mM Tris-HCl, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% Triton X-100, protease inhibitor cocktail tablet (Complete EDTA-free; catalog #11873580001, Roche) and 1 mM phenylmethanelsulfonyl fluoride (catalog #P7626, Sigma-Aldrich)] and centrifuged at 17,900 g at 4°C.and centrifuged at 17,900 g at 4°C. ... The resultant soluble and insoluble fractions were immunoblotted as described previously (Sugiura et al., 2015), and primary antibodies against FLAG (1:800; catalog #F3165, Sigma-Aldrich), Rheb1 [1:1000; catalog #MAB3426, R&D Systems (recognizing whole Rheb); 1:1000; catalog #13879; Cell Signaling Technology (preferably recognizing unfarnesylated Rheb)], a/b -tubulin (1:1000; catalog #2148, Cell Signaling Technology), phospho (p)-ERK1/2 (Thr202/Tyr204; 1:1000; catalog #9101, Cell Signaling Technology), ERK1/2 (1:1000; catalog #9102, Cell Signaling Technology), syntenin1 (1:400; catalog #sc-100336, Santa Cruz Biotechnology), p-S6 (Ser240/244; 1:1000; catalog #5364, Cell Signaling Technology), and S6 (1:1000; catalog #2317, Cell Signaling Technology), and HRP-conjugated secondary antibodies (1:6000; catalog #170–6515, BIO-RAD) were used.. Quantitative analysis of dendritic arc immunofluorescence.Quantitative analysis of dendritic arc immunofluorescence.

    In Vivo:

    Article Title: A Farnesyltransferase Inhibitor Restores Cognitive Deficits in<i>Tsc2</i><sup>+/–</sup>Mice through Inhibition of Rheb1
    Article Snippet: HEK293 T cells, cultured hippocampal neurons, and mouse brains and livers were homogenized [40 mM Tris-HCl, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% Triton X-100, protease inhibitor cocktail tablet (Complete EDTA-free; catalog #11873580001, Roche) and 1 mM phenylmethanelsulfonyl fluoride (catalog #P7626, Sigma-Aldrich)] and centrifuged at 17,900 g at 4°C.and centrifuged at 17,900 g at 4°C. ... The resultant soluble and insoluble fractions were immunoblotted as described previously (Sugiura et al., 2015), and primary antibodies against FLAG (1:800; catalog #F3165, Sigma-Aldrich), Rheb1 [1:1000; catalog #MAB3426, R&D Systems (recognizing whole Rheb); 1:1000; catalog #13879; Cell Signaling Technology (preferably recognizing unfarnesylated Rheb)], a/b -tubulin (1:1000; catalog #2148, Cell Signaling Technology), phospho (p)-ERK1/2 (Thr202/Tyr204; 1:1000; catalog #9101, Cell Signaling Technology), ERK1/2 (1:1000; catalog #9102, Cell Signaling Technology), syntenin1 (1:400; catalog #sc-100336, Santa Cruz Biotechnology), p-S6 (Ser240/244; 1:1000; catalog #5364, Cell Signaling Technology), and S6 (1:1000; catalog #2317, Cell Signaling Technology), and HRP-conjugated secondary antibodies (1:6000; catalog #170–6515, BIO-RAD) were used.. Quantitative analysis of dendritic arc immunofluorescence.Quantitative analysis of dendritic arc immunofluorescence.

    Phospho-proteomics:

    Article Title: A Farnesyltransferase Inhibitor Restores Cognitive Deficits in<i>Tsc2</i><sup>+/–</sup>Mice through Inhibition of Rheb1
    Article Snippet: HEK293 T cells, cultured hippocampal neurons, and mouse brains and livers were homogenized [40 mM Tris-HCl, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% Triton X-100, protease inhibitor cocktail tablet (Complete EDTA-free; catalog #11873580001, Roche) and 1 mM phenylmethanelsulfonyl fluoride (catalog #P7626, Sigma-Aldrich)] and centrifuged at 17,900 g at 4°C.and centrifuged at 17,900 g at 4°C. ... The resultant soluble and insoluble fractions were immunoblotted as described previously (Sugiura et al., 2015), and primary antibodies against FLAG (1:800; catalog #F3165, Sigma-Aldrich), Rheb1 [1:1000; catalog #MAB3426, R&D Systems (recognizing whole Rheb); 1:1000; catalog #13879; Cell Signaling Technology (preferably recognizing unfarnesylated Rheb)], a/b -tubulin (1:1000; catalog #2148, Cell Signaling Technology), phospho (p)-ERK1/2 (Thr202/Tyr204; 1:1000; catalog #9101, Cell Signaling Technology), ERK1/2 (1:1000; catalog #9102, Cell Signaling Technology), syntenin1 (1:400; catalog #sc-100336, Santa Cruz Biotechnology), p-S6 (Ser240/244; 1:1000; catalog #5364, Cell Signaling Technology), and S6 (1:1000; catalog #2317, Cell Signaling Technology), and HRP-conjugated secondary antibodies (1:6000; catalog #170–6515, BIO-RAD) were used.. Quantitative analysis of dendritic arc immunofluorescence.Quantitative analysis of dendritic arc immunofluorescence.

    Staining:

    Article Title: A Farnesyltransferase Inhibitor Restores Cognitive Deficits in<i>Tsc2</i><sup>+/–</sup>Mice through Inhibition of Rheb1
    Article Snippet: HEK293 T cells, cultured hippocampal neurons, and mouse brains and livers were homogenized [40 mM Tris-HCl, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% Triton X-100, protease inhibitor cocktail tablet (Complete EDTA-free; catalog #11873580001, Roche) and 1 mM phenylmethanelsulfonyl fluoride (catalog #P7626, Sigma-Aldrich)] and centrifuged at 17,900 g at 4°C.and centrifuged at 17,900 g at 4°C. ... The resultant soluble and insoluble fractions were immunoblotted as described previously (Sugiura et al., 2015), and primary antibodies against FLAG (1:800; catalog #F3165, Sigma-Aldrich), Rheb1 [1:1000; catalog #MAB3426, R&D Systems (recognizing whole Rheb); 1:1000; catalog #13879; Cell Signaling Technology (preferably recognizing unfarnesylated Rheb)], a/b -tubulin (1:1000; catalog #2148, Cell Signaling Technology), phospho (p)-ERK1/2 (Thr202/Tyr204; 1:1000; catalog #9101, Cell Signaling Technology), ERK1/2 (1:1000; catalog #9102, Cell Signaling Technology), syntenin1 (1:400; catalog #sc-100336, Santa Cruz Biotechnology), p-S6 (Ser240/244; 1:1000; catalog #5364, Cell Signaling Technology), and S6 (1:1000; catalog #2317, Cell Signaling Technology), and HRP-conjugated secondary antibodies (1:6000; catalog #170–6515, BIO-RAD) were used.. Quantitative analysis of dendritic arc immunofluorescence.Quantitative analysis of dendritic arc immunofluorescence.

    Labeling:

    Article Title: A Farnesyltransferase Inhibitor Restores Cognitive Deficits in<i>Tsc2</i><sup>+/–</sup>Mice through Inhibition of Rheb1
    Article Snippet: HEK293 T cells, cultured hippocampal neurons, and mouse brains and livers were homogenized [40 mM Tris-HCl, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% Triton X-100, protease inhibitor cocktail tablet (Complete EDTA-free; catalog #11873580001, Roche) and 1 mM phenylmethanelsulfonyl fluoride (catalog #P7626, Sigma-Aldrich)] and centrifuged at 17,900 g at 4°C.and centrifuged at 17,900 g at 4°C. ... The resultant soluble and insoluble fractions were immunoblotted as described previously (Sugiura et al., 2015), and primary antibodies against FLAG (1:800; catalog #F3165, Sigma-Aldrich), Rheb1 [1:1000; catalog #MAB3426, R&D Systems (recognizing whole Rheb); 1:1000; catalog #13879; Cell Signaling Technology (preferably recognizing unfarnesylated Rheb)], a/b -tubulin (1:1000; catalog #2148, Cell Signaling Technology), phospho (p)-ERK1/2 (Thr202/Tyr204; 1:1000; catalog #9101, Cell Signaling Technology), ERK1/2 (1:1000; catalog #9102, Cell Signaling Technology), syntenin1 (1:400; catalog #sc-100336, Santa Cruz Biotechnology), p-S6 (Ser240/244; 1:1000; catalog #5364, Cell Signaling Technology), and S6 (1:1000; catalog #2317, Cell Signaling Technology), and HRP-conjugated secondary antibodies (1:6000; catalog #170–6515, BIO-RAD) were used.. Quantitative analysis of dendritic arc immunofluorescence.Quantitative analysis of dendritic arc immunofluorescence.

    Hot Plate Test:

    Article Title: A Farnesyltransferase Inhibitor Restores Cognitive Deficits in<i>Tsc2</i><sup>+/–</sup>Mice through Inhibition of Rheb1
    Article Snippet: HEK293 T cells, cultured hippocampal neurons, and mouse brains and livers were homogenized [40 mM Tris-HCl, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% Triton X-100, protease inhibitor cocktail tablet (Complete EDTA-free; catalog #11873580001, Roche) and 1 mM phenylmethanelsulfonyl fluoride (catalog #P7626, Sigma-Aldrich)] and centrifuged at 17,900 g at 4°C.and centrifuged at 17,900 g at 4°C. ... The resultant soluble and insoluble fractions were immunoblotted as described previously (Sugiura et al., 2015), and primary antibodies against FLAG (1:800; catalog #F3165, Sigma-Aldrich), Rheb1 [1:1000; catalog #MAB3426, R&D Systems (recognizing whole Rheb); 1:1000; catalog #13879; Cell Signaling Technology (preferably recognizing unfarnesylated Rheb)], a/b -tubulin (1:1000; catalog #2148, Cell Signaling Technology), phospho (p)-ERK1/2 (Thr202/Tyr204; 1:1000; catalog #9101, Cell Signaling Technology), ERK1/2 (1:1000; catalog #9102, Cell Signaling Technology), syntenin1 (1:400; catalog #sc-100336, Santa Cruz Biotechnology), p-S6 (Ser240/244; 1:1000; catalog #5364, Cell Signaling Technology), and S6 (1:1000; catalog #2317, Cell Signaling Technology), and HRP-conjugated secondary antibodies (1:6000; catalog #170–6515, BIO-RAD) were used.. Quantitative analysis of dendritic arc immunofluorescence.Quantitative analysis of dendritic arc immunofluorescence.



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    <t>Rheb1</t> upregulates β cell mass and mTORC1 signaling. A: Rheb1 expression in the islets isolated from people with different age; B: Immunofluorescence staining of Rheb1 (red) and insulin (as a β-cell marker; green) in pancreatic sections of 8-week-old male pRheb1KO mice ( n = 3) and wild-type (WT) mice ( n = 3). Scale bars: 10 μm; C and D: Average β-cell mass and β-cell size in pancreatic sections of 2-month-old male pRheb1KO ( n = 4) and WT mice ( n = 5) subjected to insulin staining; E: Anti-apoptotic genes from RNA-seq of islets of bRheb1KO mice ( n = 3) and WT mice ( n = 3), of which is reanalyzed from our previous RNA-seq data. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01. WT: Wild-type.
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    <t>Rheb1</t> upregulates β cell mass and mTORC1 signaling. A: Rheb1 expression in the islets isolated from people with different age; B: Immunofluorescence staining of Rheb1 (red) and insulin (as a β-cell marker; green) in pancreatic sections of 8-week-old male pRheb1KO mice ( n = 3) and wild-type (WT) mice ( n = 3). Scale bars: 10 μm; C and D: Average β-cell mass and β-cell size in pancreatic sections of 2-month-old male pRheb1KO ( n = 4) and WT mice ( n = 5) subjected to insulin staining; E: Anti-apoptotic genes from RNA-seq of islets of bRheb1KO mice ( n = 3) and WT mice ( n = 3), of which is reanalyzed from our previous RNA-seq data. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01. WT: Wild-type.
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    A) Immunofluorescent labeling of endogenous CaMKIIα and the GLuc-SERCaMP in rat primay neurons transduced with AAV1 CaMKIIα GLuc-SERCaMP. B) Extracellular (n=5-6/group) and C, intracellular levels of GLuc-SERCaMP following treatment with following thapsigargin (Tg) (100 nM) (n=4/group). D) Intracellular levels of GLuc-SERCaMP 24 h following treatment with glutamate (n=3/group). E) Correlation between GLuc-SERCaMP secretion and viability glutamate treatment (n=3/group, Pearson correlation). F) Intracellular levels of GLuc-SERCaMP 24 h following treatment with kainic acid (n=3/group). G) Correlation between GLuc-SERCaMP secretion and viability kainic acid treatment (n=3/group, Pearson correlation). Data are shown as mean + SEM. *p<.05, **p<.01 vs. vehicle. *p<.05, ***p<.0005, ****p<.0001 vs. vehicle. 40X magnification, scale bars = 75 µm.
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    A) Immunofluorescent labeling of endogenous CaMKIIα and the GLuc-SERCaMP in rat primay neurons transduced with AAV1 CaMKIIα GLuc-SERCaMP. B) Extracellular (n=5-6/group) and C, intracellular levels of GLuc-SERCaMP following treatment with following thapsigargin (Tg) (100 nM) (n=4/group). D) Intracellular levels of GLuc-SERCaMP 24 h following treatment with glutamate (n=3/group). E) Correlation between GLuc-SERCaMP secretion and viability glutamate treatment (n=3/group, Pearson correlation). F) Intracellular levels of GLuc-SERCaMP 24 h following treatment with kainic acid (n=3/group). G) Correlation between GLuc-SERCaMP secretion and viability kainic acid treatment (n=3/group, Pearson correlation). Data are shown as mean + SEM. *p<.05, **p<.01 vs. vehicle. *p<.05, ***p<.0005, ****p<.0001 vs. vehicle. 40X magnification, scale bars = 75 µm.
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    A) Immunofluorescent labeling of endogenous CaMKIIα and the GLuc-SERCaMP in rat primay neurons transduced with AAV1 CaMKIIα GLuc-SERCaMP. B) Extracellular (n=5-6/group) and C, intracellular levels of GLuc-SERCaMP following treatment with following thapsigargin (Tg) (100 nM) (n=4/group). D) Intracellular levels of GLuc-SERCaMP 24 h following treatment with glutamate (n=3/group). E) Correlation between GLuc-SERCaMP secretion and viability glutamate treatment (n=3/group, Pearson correlation). F) Intracellular levels of GLuc-SERCaMP 24 h following treatment with kainic acid (n=3/group). G) Correlation between GLuc-SERCaMP secretion and viability kainic acid treatment (n=3/group, Pearson correlation). Data are shown as mean + SEM. *p<.05, **p<.01 vs. vehicle. *p<.05, ***p<.0005, ****p<.0001 vs. vehicle. 40X magnification, scale bars = 75 µm.
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    A) Immunofluorescent labeling of endogenous CaMKIIα and the GLuc-SERCaMP in rat primay neurons transduced with AAV1 CaMKIIα GLuc-SERCaMP. B) Extracellular (n=5-6/group) and C, intracellular levels of GLuc-SERCaMP following treatment with following thapsigargin (Tg) (100 nM) (n=4/group). D) Intracellular levels of GLuc-SERCaMP 24 h following treatment with glutamate (n=3/group). E) Correlation between GLuc-SERCaMP secretion and viability glutamate treatment (n=3/group, Pearson correlation). F) Intracellular levels of GLuc-SERCaMP 24 h following treatment with kainic acid (n=3/group). G) Correlation between GLuc-SERCaMP secretion and viability kainic acid treatment (n=3/group, Pearson correlation). Data are shown as mean + SEM. *p<.05, **p<.01 vs. vehicle. *p<.05, ***p<.0005, ****p<.0001 vs. vehicle. 40X magnification, scale bars = 75 µm.
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    Image Search Results


    Rheb1 upregulates β cell mass and mTORC1 signaling. A: Rheb1 expression in the islets isolated from people with different age; B: Immunofluorescence staining of Rheb1 (red) and insulin (as a β-cell marker; green) in pancreatic sections of 8-week-old male pRheb1KO mice ( n = 3) and wild-type (WT) mice ( n = 3). Scale bars: 10 μm; C and D: Average β-cell mass and β-cell size in pancreatic sections of 2-month-old male pRheb1KO ( n = 4) and WT mice ( n = 5) subjected to insulin staining; E: Anti-apoptotic genes from RNA-seq of islets of bRheb1KO mice ( n = 3) and WT mice ( n = 3), of which is reanalyzed from our previous RNA-seq data. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01. WT: Wild-type.

    Journal: World Journal of Diabetes

    Article Title: Ras homolog enriched in brain 1 regulates β cell mass and β cell function via mTORC1/AMPK/Notch1 pathways

    doi: 10.4239/wjd.v16.i6.104973

    Figure Lengend Snippet: Rheb1 upregulates β cell mass and mTORC1 signaling. A: Rheb1 expression in the islets isolated from people with different age; B: Immunofluorescence staining of Rheb1 (red) and insulin (as a β-cell marker; green) in pancreatic sections of 8-week-old male pRheb1KO mice ( n = 3) and wild-type (WT) mice ( n = 3). Scale bars: 10 μm; C and D: Average β-cell mass and β-cell size in pancreatic sections of 2-month-old male pRheb1KO ( n = 4) and WT mice ( n = 5) subjected to insulin staining; E: Anti-apoptotic genes from RNA-seq of islets of bRheb1KO mice ( n = 3) and WT mice ( n = 3), of which is reanalyzed from our previous RNA-seq data. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01. WT: Wild-type.

    Article Snippet: Islets or cells were maintained in antibiotic-free medium for 24 hours before transfection with either Rheb1 siRNA (RiboBio; siG161129043512) or scrambled siRNA (RiboBio) at a final concentration of 25 nM.

    Techniques: Expressing, Isolation, Immunofluorescence, Staining, Marker, RNA Sequencing

    Rheb1 activates mTORC1 signaling in islets. A: The expression of mTORC1 signaling and its downstream in the islets isolated from pRheb1KO ( n = 2) and Flox ( n = 2) with or without glucose stimulation; B: The expression of mTOC1 signaling and its downstream in the islets isolated from pRheb1KO ( n = 3) and Flox ( n = 3) with or without glucose and insulin stimulation; C: Western blot analysis of C57 islets treated with GFP ( n = 3) or Rheb1OE ( n = 3) adenovirus for 48 hours with or without glucose and insulin stimulation. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01. NC: Negative control.

    Journal: World Journal of Diabetes

    Article Title: Ras homolog enriched in brain 1 regulates β cell mass and β cell function via mTORC1/AMPK/Notch1 pathways

    doi: 10.4239/wjd.v16.i6.104973

    Figure Lengend Snippet: Rheb1 activates mTORC1 signaling in islets. A: The expression of mTORC1 signaling and its downstream in the islets isolated from pRheb1KO ( n = 2) and Flox ( n = 2) with or without glucose stimulation; B: The expression of mTOC1 signaling and its downstream in the islets isolated from pRheb1KO ( n = 3) and Flox ( n = 3) with or without glucose and insulin stimulation; C: Western blot analysis of C57 islets treated with GFP ( n = 3) or Rheb1OE ( n = 3) adenovirus for 48 hours with or without glucose and insulin stimulation. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01. NC: Negative control.

    Article Snippet: Islets or cells were maintained in antibiotic-free medium for 24 hours before transfection with either Rheb1 siRNA (RiboBio; siG161129043512) or scrambled siRNA (RiboBio) at a final concentration of 25 nM.

    Techniques: Expressing, Isolation, Western Blot, Negative Control

    Rheb1 promotes β cell proliferation only partly depend on the mTORC1. A: The population of 5-ethy-nyl-2’-deoxyuridine + cells of MIN6 cells treated with PWPI or Rheb1OE lentivirus with or without rapamycin treatment; B: The population of Ki67+ cells of insulin+ islet treated with GFP or Rheb1OE adenovirus with or without rapamycin treatment; C: Western blot analysis of C57 mice islets treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin treatment ( n = 3/group). All data are represented as the mean ± SD, a P < 0.05, b P < 0.001. NC: Negative control; Edu: 5-ethy-nyl-2’-deoxyuridine.

    Journal: World Journal of Diabetes

    Article Title: Ras homolog enriched in brain 1 regulates β cell mass and β cell function via mTORC1/AMPK/Notch1 pathways

    doi: 10.4239/wjd.v16.i6.104973

    Figure Lengend Snippet: Rheb1 promotes β cell proliferation only partly depend on the mTORC1. A: The population of 5-ethy-nyl-2’-deoxyuridine + cells of MIN6 cells treated with PWPI or Rheb1OE lentivirus with or without rapamycin treatment; B: The population of Ki67+ cells of insulin+ islet treated with GFP or Rheb1OE adenovirus with or without rapamycin treatment; C: Western blot analysis of C57 mice islets treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin treatment ( n = 3/group). All data are represented as the mean ± SD, a P < 0.05, b P < 0.001. NC: Negative control; Edu: 5-ethy-nyl-2’-deoxyuridine.

    Article Snippet: Islets or cells were maintained in antibiotic-free medium for 24 hours before transfection with either Rheb1 siRNA (RiboBio; siG161129043512) or scrambled siRNA (RiboBio) at a final concentration of 25 nM.

    Techniques: Western Blot, Negative Control

    Rheb1 regulates β cell proliferation via mTORC1 and AMPK signaling simultaneously. A: The expression of AMPK signaling and its downstream in the MIN6 cell treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin treatment; B: The population of 5-ethy-nyl-2’-deoxyuridine + cells of MIN6 cell treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin and AICAR treatment; C: The population of Ki67+ cells of insulin+ islet treated with GFP or Rheb1OE adenovirus with or without rapamycin and AICAR treatment; D: Western blot analysis of MIN6 cell treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin and AICAR treatment; E: Cell vitality analysis of MIN6 cell treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin and AICAR treatment. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01, c P < 0.001.

    Journal: World Journal of Diabetes

    Article Title: Ras homolog enriched in brain 1 regulates β cell mass and β cell function via mTORC1/AMPK/Notch1 pathways

    doi: 10.4239/wjd.v16.i6.104973

    Figure Lengend Snippet: Rheb1 regulates β cell proliferation via mTORC1 and AMPK signaling simultaneously. A: The expression of AMPK signaling and its downstream in the MIN6 cell treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin treatment; B: The population of 5-ethy-nyl-2’-deoxyuridine + cells of MIN6 cell treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin and AICAR treatment; C: The population of Ki67+ cells of insulin+ islet treated with GFP or Rheb1OE adenovirus with or without rapamycin and AICAR treatment; D: Western blot analysis of MIN6 cell treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin and AICAR treatment; E: Cell vitality analysis of MIN6 cell treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin and AICAR treatment. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01, c P < 0.001.

    Article Snippet: Islets or cells were maintained in antibiotic-free medium for 24 hours before transfection with either Rheb1 siRNA (RiboBio; siG161129043512) or scrambled siRNA (RiboBio) at a final concentration of 25 nM.

    Techniques: Expressing, Western Blot

    Rheb1 keeps β-cell identity balance through mTORC1 and Notch1 signaling. A: Average α-cell proportion in pancreatic sections of 2-month-old male bRheb1KO and WT mice subjected to glucagon staining ( n = 4/group); B and C: The mRNA (B) and protein (C) levels of β-cell dedifferentiation related markers in MIN6 cell infected with GFP or Rheb1 adenovirus; D: Western blot analysis of MIN6 cell treated with a Rheb1-specific siRNA or a siRNA control for 48 hours; E: The mRNA levels of Notch pathway in MIN6 cells infected with GFP or Rheb1OE adenovirus for 48 hours; F: Western blot analysis of MIN6 cell infected with GFP or Rheb1OE adenovirus, treated with different levels of rapamycin and DAPT. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01, c P < 0.001. NC: Negative control; WT: Wild-type.

    Journal: World Journal of Diabetes

    Article Title: Ras homolog enriched in brain 1 regulates β cell mass and β cell function via mTORC1/AMPK/Notch1 pathways

    doi: 10.4239/wjd.v16.i6.104973

    Figure Lengend Snippet: Rheb1 keeps β-cell identity balance through mTORC1 and Notch1 signaling. A: Average α-cell proportion in pancreatic sections of 2-month-old male bRheb1KO and WT mice subjected to glucagon staining ( n = 4/group); B and C: The mRNA (B) and protein (C) levels of β-cell dedifferentiation related markers in MIN6 cell infected with GFP or Rheb1 adenovirus; D: Western blot analysis of MIN6 cell treated with a Rheb1-specific siRNA or a siRNA control for 48 hours; E: The mRNA levels of Notch pathway in MIN6 cells infected with GFP or Rheb1OE adenovirus for 48 hours; F: Western blot analysis of MIN6 cell infected with GFP or Rheb1OE adenovirus, treated with different levels of rapamycin and DAPT. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01, c P < 0.001. NC: Negative control; WT: Wild-type.

    Article Snippet: Islets or cells were maintained in antibiotic-free medium for 24 hours before transfection with either Rheb1 siRNA (RiboBio; siG161129043512) or scrambled siRNA (RiboBio) at a final concentration of 25 nM.

    Techniques: Staining, Infection, Western Blot, Control, Negative Control

    Rheb1 regulates transcription factor HNF4α. A: Protein expression level of HNF4α was increased in Rheb1 overexpressed MIN6 cells ( n = 3); B: Protein expression level of HNF4α was decreased in Rheb1 knockout MIN6 cells; C: The interaction between Rheb1 and HNF4α were analyzed by co-IP in MIN6 cells; D: After overexpression Rheb1 in human islets for 48 hours, immunofluorescence staining of Rheb1 and HNF4α in human islets. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01. NC: Negative control.

    Journal: World Journal of Diabetes

    Article Title: Ras homolog enriched in brain 1 regulates β cell mass and β cell function via mTORC1/AMPK/Notch1 pathways

    doi: 10.4239/wjd.v16.i6.104973

    Figure Lengend Snippet: Rheb1 regulates transcription factor HNF4α. A: Protein expression level of HNF4α was increased in Rheb1 overexpressed MIN6 cells ( n = 3); B: Protein expression level of HNF4α was decreased in Rheb1 knockout MIN6 cells; C: The interaction between Rheb1 and HNF4α were analyzed by co-IP in MIN6 cells; D: After overexpression Rheb1 in human islets for 48 hours, immunofluorescence staining of Rheb1 and HNF4α in human islets. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01. NC: Negative control.

    Article Snippet: Islets or cells were maintained in antibiotic-free medium for 24 hours before transfection with either Rheb1 siRNA (RiboBio; siG161129043512) or scrambled siRNA (RiboBio) at a final concentration of 25 nM.

    Techniques: Expressing, Knock-Out, Co-Immunoprecipitation Assay, Over Expression, Immunofluorescence, Staining, Negative Control

    Rheb1 upregulates β cell mass and mTORC1 signaling. A: Rheb1 expression in the islets isolated from people with different age; B: Immunofluorescence staining of Rheb1 (red) and insulin (as a β-cell marker; green) in pancreatic sections of 8-week-old male pRheb1KO mice ( n = 3) and wild-type (WT) mice ( n = 3). Scale bars: 10 μm; C and D: Average β-cell mass and β-cell size in pancreatic sections of 2-month-old male pRheb1KO ( n = 4) and WT mice ( n = 5) subjected to insulin staining; E: Anti-apoptotic genes from RNA-seq of islets of bRheb1KO mice ( n = 3) and WT mice ( n = 3), of which is reanalyzed from our previous RNA-seq data. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01. WT: Wild-type.

    Journal: World Journal of Diabetes

    Article Title: Ras homolog enriched in brain 1 regulates β cell mass and β cell function via mTORC1/AMPK/Notch1 pathways

    doi: 10.4239/wjd.v16.i6.104973

    Figure Lengend Snippet: Rheb1 upregulates β cell mass and mTORC1 signaling. A: Rheb1 expression in the islets isolated from people with different age; B: Immunofluorescence staining of Rheb1 (red) and insulin (as a β-cell marker; green) in pancreatic sections of 8-week-old male pRheb1KO mice ( n = 3) and wild-type (WT) mice ( n = 3). Scale bars: 10 μm; C and D: Average β-cell mass and β-cell size in pancreatic sections of 2-month-old male pRheb1KO ( n = 4) and WT mice ( n = 5) subjected to insulin staining; E: Anti-apoptotic genes from RNA-seq of islets of bRheb1KO mice ( n = 3) and WT mice ( n = 3), of which is reanalyzed from our previous RNA-seq data. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01. WT: Wild-type.

    Article Snippet: Specific antibodies were used to detect Rheb1 (CST 13879S), Cyclin D1 (CST 5506S), Cyclin D2 (CST 3741S), phosphorylated S6 at Ser235 (CST 5364S), phosphorylated 4EBP1 at Thr37/46 (CST 9459S), total S6 (CST 2217S), and total 4EBP1 (CST 2855S).

    Techniques: Expressing, Isolation, Immunofluorescence, Staining, Marker, RNA Sequencing

    Rheb1 activates mTORC1 signaling in islets. A: The expression of mTORC1 signaling and its downstream in the islets isolated from pRheb1KO ( n = 2) and Flox ( n = 2) with or without glucose stimulation; B: The expression of mTOC1 signaling and its downstream in the islets isolated from pRheb1KO ( n = 3) and Flox ( n = 3) with or without glucose and insulin stimulation; C: Western blot analysis of C57 islets treated with GFP ( n = 3) or Rheb1OE ( n = 3) adenovirus for 48 hours with or without glucose and insulin stimulation. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01. NC: Negative control.

    Journal: World Journal of Diabetes

    Article Title: Ras homolog enriched in brain 1 regulates β cell mass and β cell function via mTORC1/AMPK/Notch1 pathways

    doi: 10.4239/wjd.v16.i6.104973

    Figure Lengend Snippet: Rheb1 activates mTORC1 signaling in islets. A: The expression of mTORC1 signaling and its downstream in the islets isolated from pRheb1KO ( n = 2) and Flox ( n = 2) with or without glucose stimulation; B: The expression of mTOC1 signaling and its downstream in the islets isolated from pRheb1KO ( n = 3) and Flox ( n = 3) with or without glucose and insulin stimulation; C: Western blot analysis of C57 islets treated with GFP ( n = 3) or Rheb1OE ( n = 3) adenovirus for 48 hours with or without glucose and insulin stimulation. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01. NC: Negative control.

    Article Snippet: Specific antibodies were used to detect Rheb1 (CST 13879S), Cyclin D1 (CST 5506S), Cyclin D2 (CST 3741S), phosphorylated S6 at Ser235 (CST 5364S), phosphorylated 4EBP1 at Thr37/46 (CST 9459S), total S6 (CST 2217S), and total 4EBP1 (CST 2855S).

    Techniques: Expressing, Isolation, Western Blot, Negative Control

    Rheb1 promotes β cell proliferation only partly depend on the mTORC1. A: The population of 5-ethy-nyl-2’-deoxyuridine + cells of MIN6 cells treated with PWPI or Rheb1OE lentivirus with or without rapamycin treatment; B: The population of Ki67+ cells of insulin+ islet treated with GFP or Rheb1OE adenovirus with or without rapamycin treatment; C: Western blot analysis of C57 mice islets treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin treatment ( n = 3/group). All data are represented as the mean ± SD, a P < 0.05, b P < 0.001. NC: Negative control; Edu: 5-ethy-nyl-2’-deoxyuridine.

    Journal: World Journal of Diabetes

    Article Title: Ras homolog enriched in brain 1 regulates β cell mass and β cell function via mTORC1/AMPK/Notch1 pathways

    doi: 10.4239/wjd.v16.i6.104973

    Figure Lengend Snippet: Rheb1 promotes β cell proliferation only partly depend on the mTORC1. A: The population of 5-ethy-nyl-2’-deoxyuridine + cells of MIN6 cells treated with PWPI or Rheb1OE lentivirus with or without rapamycin treatment; B: The population of Ki67+ cells of insulin+ islet treated with GFP or Rheb1OE adenovirus with or without rapamycin treatment; C: Western blot analysis of C57 mice islets treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin treatment ( n = 3/group). All data are represented as the mean ± SD, a P < 0.05, b P < 0.001. NC: Negative control; Edu: 5-ethy-nyl-2’-deoxyuridine.

    Article Snippet: Specific antibodies were used to detect Rheb1 (CST 13879S), Cyclin D1 (CST 5506S), Cyclin D2 (CST 3741S), phosphorylated S6 at Ser235 (CST 5364S), phosphorylated 4EBP1 at Thr37/46 (CST 9459S), total S6 (CST 2217S), and total 4EBP1 (CST 2855S).

    Techniques: Western Blot, Negative Control

    Rheb1 regulates β cell proliferation via mTORC1 and AMPK signaling simultaneously. A: The expression of AMPK signaling and its downstream in the MIN6 cell treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin treatment; B: The population of 5-ethy-nyl-2’-deoxyuridine + cells of MIN6 cell treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin and AICAR treatment; C: The population of Ki67+ cells of insulin+ islet treated with GFP or Rheb1OE adenovirus with or without rapamycin and AICAR treatment; D: Western blot analysis of MIN6 cell treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin and AICAR treatment; E: Cell vitality analysis of MIN6 cell treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin and AICAR treatment. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01, c P < 0.001.

    Journal: World Journal of Diabetes

    Article Title: Ras homolog enriched in brain 1 regulates β cell mass and β cell function via mTORC1/AMPK/Notch1 pathways

    doi: 10.4239/wjd.v16.i6.104973

    Figure Lengend Snippet: Rheb1 regulates β cell proliferation via mTORC1 and AMPK signaling simultaneously. A: The expression of AMPK signaling and its downstream in the MIN6 cell treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin treatment; B: The population of 5-ethy-nyl-2’-deoxyuridine + cells of MIN6 cell treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin and AICAR treatment; C: The population of Ki67+ cells of insulin+ islet treated with GFP or Rheb1OE adenovirus with or without rapamycin and AICAR treatment; D: Western blot analysis of MIN6 cell treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin and AICAR treatment; E: Cell vitality analysis of MIN6 cell treated with GFP or Rheb1OE adenovirus for 48 hours with or without rapamycin and AICAR treatment. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01, c P < 0.001.

    Article Snippet: Specific antibodies were used to detect Rheb1 (CST 13879S), Cyclin D1 (CST 5506S), Cyclin D2 (CST 3741S), phosphorylated S6 at Ser235 (CST 5364S), phosphorylated 4EBP1 at Thr37/46 (CST 9459S), total S6 (CST 2217S), and total 4EBP1 (CST 2855S).

    Techniques: Expressing, Western Blot

    Rheb1 keeps β-cell identity balance through mTORC1 and Notch1 signaling. A: Average α-cell proportion in pancreatic sections of 2-month-old male bRheb1KO and WT mice subjected to glucagon staining ( n = 4/group); B and C: The mRNA (B) and protein (C) levels of β-cell dedifferentiation related markers in MIN6 cell infected with GFP or Rheb1 adenovirus; D: Western blot analysis of MIN6 cell treated with a Rheb1-specific siRNA or a siRNA control for 48 hours; E: The mRNA levels of Notch pathway in MIN6 cells infected with GFP or Rheb1OE adenovirus for 48 hours; F: Western blot analysis of MIN6 cell infected with GFP or Rheb1OE adenovirus, treated with different levels of rapamycin and DAPT. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01, c P < 0.001. NC: Negative control; WT: Wild-type.

    Journal: World Journal of Diabetes

    Article Title: Ras homolog enriched in brain 1 regulates β cell mass and β cell function via mTORC1/AMPK/Notch1 pathways

    doi: 10.4239/wjd.v16.i6.104973

    Figure Lengend Snippet: Rheb1 keeps β-cell identity balance through mTORC1 and Notch1 signaling. A: Average α-cell proportion in pancreatic sections of 2-month-old male bRheb1KO and WT mice subjected to glucagon staining ( n = 4/group); B and C: The mRNA (B) and protein (C) levels of β-cell dedifferentiation related markers in MIN6 cell infected with GFP or Rheb1 adenovirus; D: Western blot analysis of MIN6 cell treated with a Rheb1-specific siRNA or a siRNA control for 48 hours; E: The mRNA levels of Notch pathway in MIN6 cells infected with GFP or Rheb1OE adenovirus for 48 hours; F: Western blot analysis of MIN6 cell infected with GFP or Rheb1OE adenovirus, treated with different levels of rapamycin and DAPT. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01, c P < 0.001. NC: Negative control; WT: Wild-type.

    Article Snippet: Specific antibodies were used to detect Rheb1 (CST 13879S), Cyclin D1 (CST 5506S), Cyclin D2 (CST 3741S), phosphorylated S6 at Ser235 (CST 5364S), phosphorylated 4EBP1 at Thr37/46 (CST 9459S), total S6 (CST 2217S), and total 4EBP1 (CST 2855S).

    Techniques: Staining, Infection, Western Blot, Control, Negative Control

    Rheb1 regulates transcription factor HNF4α. A: Protein expression level of HNF4α was increased in Rheb1 overexpressed MIN6 cells ( n = 3); B: Protein expression level of HNF4α was decreased in Rheb1 knockout MIN6 cells; C: The interaction between Rheb1 and HNF4α were analyzed by co-IP in MIN6 cells; D: After overexpression Rheb1 in human islets for 48 hours, immunofluorescence staining of Rheb1 and HNF4α in human islets. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01. NC: Negative control.

    Journal: World Journal of Diabetes

    Article Title: Ras homolog enriched in brain 1 regulates β cell mass and β cell function via mTORC1/AMPK/Notch1 pathways

    doi: 10.4239/wjd.v16.i6.104973

    Figure Lengend Snippet: Rheb1 regulates transcription factor HNF4α. A: Protein expression level of HNF4α was increased in Rheb1 overexpressed MIN6 cells ( n = 3); B: Protein expression level of HNF4α was decreased in Rheb1 knockout MIN6 cells; C: The interaction between Rheb1 and HNF4α were analyzed by co-IP in MIN6 cells; D: After overexpression Rheb1 in human islets for 48 hours, immunofluorescence staining of Rheb1 and HNF4α in human islets. All data are represented as the mean ± SD, a P < 0.05, b P < 0.01. NC: Negative control.

    Article Snippet: Specific antibodies were used to detect Rheb1 (CST 13879S), Cyclin D1 (CST 5506S), Cyclin D2 (CST 3741S), phosphorylated S6 at Ser235 (CST 5364S), phosphorylated 4EBP1 at Thr37/46 (CST 9459S), total S6 (CST 2217S), and total 4EBP1 (CST 2855S).

    Techniques: Expressing, Knock-Out, Co-Immunoprecipitation Assay, Over Expression, Immunofluorescence, Staining, Negative Control

    A) Immunofluorescent labeling of endogenous CaMKIIα and the GLuc-SERCaMP in rat primay neurons transduced with AAV1 CaMKIIα GLuc-SERCaMP. B) Extracellular (n=5-6/group) and C, intracellular levels of GLuc-SERCaMP following treatment with following thapsigargin (Tg) (100 nM) (n=4/group). D) Intracellular levels of GLuc-SERCaMP 24 h following treatment with glutamate (n=3/group). E) Correlation between GLuc-SERCaMP secretion and viability glutamate treatment (n=3/group, Pearson correlation). F) Intracellular levels of GLuc-SERCaMP 24 h following treatment with kainic acid (n=3/group). G) Correlation between GLuc-SERCaMP secretion and viability kainic acid treatment (n=3/group, Pearson correlation). Data are shown as mean + SEM. *p<.05, **p<.01 vs. vehicle. *p<.05, ***p<.0005, ****p<.0001 vs. vehicle. 40X magnification, scale bars = 75 µm.

    Journal: bioRxiv

    Article Title: Excitotoxic glutamate levels cause the secretion of resident endoplasmic reticulum proteins

    doi: 10.1101/2023.09.03.556116

    Figure Lengend Snippet: A) Immunofluorescent labeling of endogenous CaMKIIα and the GLuc-SERCaMP in rat primay neurons transduced with AAV1 CaMKIIα GLuc-SERCaMP. B) Extracellular (n=5-6/group) and C, intracellular levels of GLuc-SERCaMP following treatment with following thapsigargin (Tg) (100 nM) (n=4/group). D) Intracellular levels of GLuc-SERCaMP 24 h following treatment with glutamate (n=3/group). E) Correlation between GLuc-SERCaMP secretion and viability glutamate treatment (n=3/group, Pearson correlation). F) Intracellular levels of GLuc-SERCaMP 24 h following treatment with kainic acid (n=3/group). G) Correlation between GLuc-SERCaMP secretion and viability kainic acid treatment (n=3/group, Pearson correlation). Data are shown as mean + SEM. *p<.05, **p<.01 vs. vehicle. *p<.05, ***p<.0005, ****p<.0001 vs. vehicle. 40X magnification, scale bars = 75 µm.

    Article Snippet: All plasmids were constructed using ligation-independent cloning (In-Fusion, Clontech) into the backbone of pAAV CaMKII iRFP-FLAG (Addgene 149513) after digestion with NheI and AscI restriction enzymes (New England Biolabs) and insert amplification by polymerase chain reaction.

    Techniques: Labeling, Transduction