ac p53 rabbit  (Cell Signaling Technology Inc)

Journal: Aging Cell

Article Title: Targeting IGF1 ‐Induced Cellular Senescence to Rejuvenate Hair Follicle Aging

doi: 10.1111/acel.70053

Figure Lengend Snippet: Elevated IGF‐1 expression accelerates hair follicle aging through p53‐dependent cellular senescence. (a) Protein expressions of IGF‐1 in mouse skin are assessed by immunohistochemistry (IHC) and show a significant increase in aged mice. Representative staining and average optical density (AOD) quantification were demonstrated. Data are presented as means ± SEM, Two‐tailed Student's t ‐test. (b) IHC was performed on human perineal skin to measure IGF‐1 expression in two age groups: Younger women (< 30 years) and older women (> 60 years). Representative staining and AOD quantification were demonstrated. AOD analysis was performed on a minimum of 200 cells from the dorsal skin. Data are presented as means ± SEM, Two‐tailed Student's t ‐test. (c) Kaplan–Meier plot shows the lifespan follow‐up in male IGF‐1 Tg mice ( n = 36, red) or WT littermates ( n = 19, blue). p values were determined by the log‐rank (Mantel‐Cox) test. (d and e) Clinical frailty index scores and rotarod test of 10‐month WT and IGF‐1 Tg male mice ( n = 8/group). Data were means ± SEM. Statistical analysis was performed using a two‐tailed Student's t ‐test. (f) Representative images of dorsal skin coat color from 10‐month‐old IGF‐1 Tg mice, 10‐month‐old WT mice, and 24‐month‐old WT mice are shown. The age‐dependent appearance of gray hair in mice is graphed and quantified. (g) 10‐month‐old IGF‐1 Tg male mice and age‐matched WT male mice were shaved and monitored for hair coat recovery. Hair regrowth was quantified as the percentage of back skin covered by new hairs. Data are shown as means ± SD, n = 5/group. Two‐tailed Student's t ‐test. (h) Equal amounts of total tissue lysates from the dorsal skin of two 10‐month‐old IGF‐1 Tg male mice and their WT littermates were immunoprecipitated and analyzed by Western blotting to detect p53 and p21. The immunoprecipitated lysates were normalized for total p53, and the expression level of acetylated p53 at lysine 379 (Ac‐K379‐p53) was quantified. (i) Histological examination of dorsal skin was performed on 10‐month‐old IGF‐1 Tg mice, 10‐month‐old WT mice, and 24‐month‐old WT mice. Representative IHC images and quantitative analyses of IGF‐1, p53, p16, and PAI‐1 were presented. Data are shown as means ± SEM, n = 5/group. Two‐way ANOVA with Tukey's test. Scale bar = 50 μm.

Article Snippet: Antibodies for Ac‐K379‐p53 (#2570S, WB 1:1000), γ‐H2AX (#2577, WB 1:1000, IF 1:200), and PAI‐1 (#11907, WB 1:1000, IHC 1:200) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Immunohistochemistry, Staining, Two Tailed Test, Immunoprecipitation, Western Blot

Journal: Aging Cell

Article Title: Targeting IGF1 ‐Induced Cellular Senescence to Rejuvenate Hair Follicle Aging

doi: 10.1111/acel.70053

Figure Lengend Snippet: Forced SIRT1 overexpression attenuates hair graying and hair follicle cell senescence in IGF‐1 Tg mice. (a) Representative images of age‐associated coat color from 10‐month‐old male IGF‐1 Tg mice, SIRT1 Tg mice, IGF‐1/SIRT1 double transgenic (DTg) mice, and WT littermates, showing dorsal gray hair ( n = 10/group). The percentage of mice with gray hair in each group is also presented. (b) Equal amounts of total tissue lysates from the dorsal skin of these mice were immunoprecipitated and analyzed by Western blotting with a p53‐specific antibody. The immunoprecipitated samples were normalized to total p53 levels before immunoblotting for Ac‐K379‐p53. (c) Histological examination of dorsal skin was conducted to assess IGF‐1 signaling. Representative IHC images and corresponding quantitative analyses are shown for IGF‐1, SIRT1, p53, p16, and PAI‐1 ( n = 8/group). (d–g) Representative IF images and corresponding quantitative analyses of senescence‐associated DNA damage marker γ‐H2AX, SASP factor IL‐6, HFSC marker CD34, and melanocyte stem cell marker KIT ( n = 5/group). Data are presented as means ± SEM; two‐way ANOVA with Tukey's test. Scale bar = 50 μm.

Article Snippet: Antibodies for Ac‐K379‐p53 (#2570S, WB 1:1000), γ‐H2AX (#2577, WB 1:1000, IF 1:200), and PAI‐1 (#11907, WB 1:1000, IHC 1:200) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Over Expression, Transgenic Assay, Immunoprecipitation, Western Blot, Marker

Journal: Aging Cell

Article Title: Targeting IGF1 ‐Induced Cellular Senescence to Rejuvenate Hair Follicle Aging

doi: 10.1111/acel.70053

Figure Lengend Snippet: Dietary Restriction targeting IGF‐1 alleviates hair graying and HFSC senescence. (a) Four‐month‐old male IGF‐1 Tg mice and WT littermates were fed either regular chow or subjected to a 30% dietary restriction ( n = 11/group) for 6 months. Dorsal coat color and hair graying were assessed, followed by IHC and IF analyses of hair follicles. (b) Representative images of dorsal coat color and percentages of mice with gray hair in each group were analyzed. (c) Representative IHC images and corresponding quantitative analyses of IGF‐1, SIRT1, p53, p16, and PAI‐1 ( n = 7/group). (d–g) Representative IF images and corresponding quantitative analyses of the senescence‐associated DNA damage marker γ‐H2AX (d). SASP factor IL‐6 (e), hair follicle stem cell marker CD34 (f), and melanocyte stem cell marker KIT (g) ( n = 5/group). Data were means ± SEM, Two‐way ANOVA with Tukey's test. Scale bar = 50 μm. (h–j) 4‐month‐old male IGF‐1 Tg mice and WT littermates were fed either regular chow or subjected to a 30% dietary restriction ( n = 5/group) for 20 months. Dorsal coat color and hair graying were assessed, followed by IHC and IF analyses of hair follicles (h). Representative coat color images and AOD quantification are shown ( n = 5/group) (i). Representative IHC images and corresponding quantitative analyses of p53, p16, and PAI‐1 ( n = 5/group) (j).

Article Snippet: Antibodies for Ac‐K379‐p53 (#2570S, WB 1:1000), γ‐H2AX (#2577, WB 1:1000, IF 1:200), and PAI‐1 (#11907, WB 1:1000, IHC 1:200) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Marker

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Knockout of onecut2 inhibits proliferation and promotes apoptosis of tumor cells through SKP2-mediated p53 acetylation in hepatocellular carcinoma

doi: 10.1007/s00018-024-05518-3

Figure Lengend Snippet: Role of proliferation and apoptosis mediated by the overexpression of OC2 and SKP2 in B8 cells. A The proliferation of B8 cells overexpressing OC2 was analyzed via a CCK-8 assay. B Flow cytometry assay of apoptosis levels in B8 cells overexpressing OC2. C The expression of apoptosis-related factors (SKP2, PARP, Bax, Bcl2, p21 and p27) in B8 cells overexpressing OC2. D Proliferation of B8 cells overexpressing SKP2. E Flow cytometry assay of apoptosis levels in B8 cells overexpressing SKP2. F - G The expression of apoptosis-related factors in B8 cells overexpressing SKP2 was analyzed by Western blotting and qRT‒PCR. H The phosphorylation and acetylation levels of p53 were analyzed by Western blotting in B8 cells overexpressing SKP2. I Dual-luciferase reporter assays of p53 response element-related fluorescence activity in SK-Hep1 cells with OC2 and SKP2 knockdown. The “p53-RE” group consisted of SK-Hep1 cells transfected with pGPL3-Basic plasmid with p53 response element sequence inserted. J Phosphorylation and acetylation levels of p53 in B8 cells overexpressing OC2 and SKP2. The data are expressed as mean ± SD; “ns” indicates no significant difference; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The anti-Ac-p53 (2525), anti-AKT (4685), anti-angiogenin (62224), anti-Bax (41162), anti-Bcl-2 (4223), anti-CD31 (77699), anti-FGF2 (46879), anti-GAPDH (5174), anti-MAPK (4695), anti-PARP (9532), anti-PDGF-B (3169), anti-p21 (2947), anti-p27 (3686), anti-p53 (2527), anti-p-AKT (4060), anti-p-MAPK (9101), anti-p-p53 (9284), anti-TGF-β1 (3709), and anti-VEGFA (50661) antibodies were purchased from Cell Signaling Technology.

Techniques: Over Expression, CCK-8 Assay, Flow Cytometry, Expressing, Western Blot, Phospho-proteomics, Luciferase, Fluorescence, Activity Assay, Knockdown, Transfection, Plasmid Preparation, Sequencing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Knockout of onecut2 inhibits proliferation and promotes apoptosis of tumor cells through SKP2-mediated p53 acetylation in hepatocellular carcinoma

doi: 10.1007/s00018-024-05518-3

Figure Lengend Snippet: The mechanism by which OC2 regulates proliferation and apoptosis in HCC cells. OC2 promoted the transcriptional activation of SKP2, which inhibited the binding of p53 to p300 and downregulated the acetylation level of p53, thereby inhibiting the apoptosis of HCC cells. The solid line indicates direct action; the dotted line indicates indirect action; the “AC” indicates acetylation; the “P” indicates phosphorylation

Article Snippet: The anti-Ac-p53 (2525), anti-AKT (4685), anti-angiogenin (62224), anti-Bax (41162), anti-Bcl-2 (4223), anti-CD31 (77699), anti-FGF2 (46879), anti-GAPDH (5174), anti-MAPK (4695), anti-PARP (9532), anti-PDGF-B (3169), anti-p21 (2947), anti-p27 (3686), anti-p53 (2527), anti-p-AKT (4060), anti-p-MAPK (9101), anti-p-p53 (9284), anti-TGF-β1 (3709), and anti-VEGFA (50661) antibodies were purchased from Cell Signaling Technology.

Techniques: Activation Assay, Binding Assay, Phospho-proteomics