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Cell Signaling Technology Inc ki 67 d3b5
Ki 67 D3b5, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 817 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ki+67+d3b5/Ki-67+Rabbit+mAb/pmc13043174-3-2-7
Average 97 stars, based on 817 article reviews

ki 67 d3b5 - by Bioz Stars, 2026-09

97/100 stars

Images

Incubation:

Article Title: The natural product salicin alleviates osteoarthritis progression by binding to IRE1α and inhibiting endoplasmic reticulum stress through the IRE1α-IκBα-p65 signaling pathway
Article Snippet: .. Then, the sections were incubated overnight at 4 °C with primary phospho-NF-κB p65 (CST), Ki-67 (D3B5) (CST) and p-IRE1α (Zen Bio) antibodies. ..

other:

Article Title: Human receptive endometrial assembloid for deciphering the implantation window
Article Snippet: Antibody , Ki-67 (D3B5) (Rabbit monoclonal) , Cell Signaling , Cat#: 9129 RRID: AB_2687446 , IF (1:400).

Article Title: Efficient improvement of the proliferation, differentiation, and anti-arthritic capacity of mesenchymal stem cells by simply culturing on the immobilized FGF2 derived peptide, 44-ERGVVSIKGV-53.
Article Snippet: WB: Western blot Signaling Technology) and rabbit anti-Ki-67 (Cat No. 9129, Cell Signaling Technology), and the secondary antibody was DyLightTM 549 Goat Anti-Rabbit IgG (Cat No. DI-1549–1.5, Vector laboratories, Burlingame, CA, USA) and Alexa Fluor 488 goat anti-rabbit IgG (Cat No. A11008, Invitrogen) which were diluted in 3 % blocking buffer.

Immunohistochemistry:

Article Title: MDM2 E3 ligase activity is essential for p53 regulation and cell cycle integrity
Article Snippet: HA-MDM2 isoforms and FLAG-MDM4 were detected with either anti-HA (HA.11, Covance, Princeton, NJ) or anti-FLAG (Sigma, M2, F1804). .. Antibodies for immunohistochemistry includes phospho-H3 (S10) (Millipore-Sigma, Cat# 06–057) and Ki-67 (D3B5) (Cell Signaling, Cat# 12202S). ..

Article Title: Circ_0002669 promotes osteosarcoma tumorigenesis through directly binding to MYCBP and sponging miR-889-3p
Article Snippet: .. Antibodies used for IHC were as follows: Ki-67 (D3B5) (1:200, #12,202, CST), cleaved caspase 3 (35A1E) (1:100, #9664, CST) and MYCBP (2E9) (1:100, sc-517,020, Santa Cruz). .. Images were captured with an Olympus microscope (Japan).

FACS:

Article Title: Efficient improvement of the proliferation, differentiation, and anti-arthritic capacity of mesenchymal stem cells by simply culturing on the immobilized FGF2 derived peptide, 44-ERGVVSIKGV-53.
Article Snippet: .. Antibody Assay Dilution Company source Catalog Number CD34 FACS 1:200 R&D system MAB72271 CD45 PD7/26/16 + 2B11- 1:200 Invitrogen MA5-13197 CD73/NT5E 1:200 Invitrogen RG235718 CD90/Thy1 1:200 R&D system AF2067 CD105 1:200 Invitrogen MA5-11854 anti-Mouse IgG Alexa Fluor 488 1:200 Invitrogen A11059 anti-Sheep lgG PE 1:200 R&D system F0126 PCNA (D3H8P) XP Immunofluorescent staining 1:400 CST 13,110 Ki-67 (D3B5) 1:400 CST 9129 anti-rabbit IgG Alexa Fluor 488 1:200 Invitrogen A11008 anti-rabbit IgG DyLight TM 549 1:200 Vector laboratories DI-1549–1.5 P-AKT WB 1:500 CUSABIO CSB-PA008120 T-AKT 1:500 CUSABIO CSB-PA000855 P-ERK 1:1,000 CST 9101 s T-ERK 1:1,000 Assaybio B7074 T-FGFR1 1:1,000 Novus Biologicals NBP2-33784 P- FGFR1 (Tyr653, Tyr654) 1:1,000 Invitrogen 44-1140G T-FRS2 1:125 Biovision 3836–100 P-FRS2-alpha 1:1,000 CST 3864S IKKa 1:500 CST 3G12 b-actin 1:1,000 Santa cruz sc-8432 HRP linked anti-rabbit IgG 1:2,000 CST 7074 HRP linked anti-mouse IgG 1:2,000 CST 7076 FACS: Fluorescence-activated Cell Sorting. .. WB: Western blot Signaling Technology) and rabbit anti-Ki-67 (Cat No. 9129, Cell Signaling Technology), and the secondary antibody was DyLightTM 549 Goat Anti-Rabbit IgG (Cat No. DI-1549–1.5, Vector laboratories, Burlingame, CA, USA) and Alexa Fluor 488 goat anti-rabbit IgG (Cat No. A11008, Invitrogen) which were diluted in 3 % blocking buffer.

Staining:

Article Title: Efficient improvement of the proliferation, differentiation, and anti-arthritic capacity of mesenchymal stem cells by simply culturing on the immobilized FGF2 derived peptide, 44-ERGVVSIKGV-53.
Article Snippet: .. Antibody Assay Dilution Company source Catalog Number CD34 FACS 1:200 R&D system MAB72271 CD45 PD7/26/16 + 2B11- 1:200 Invitrogen MA5-13197 CD73/NT5E 1:200 Invitrogen RG235718 CD90/Thy1 1:200 R&D system AF2067 CD105 1:200 Invitrogen MA5-11854 anti-Mouse IgG Alexa Fluor 488 1:200 Invitrogen A11059 anti-Sheep lgG PE 1:200 R&D system F0126 PCNA (D3H8P) XP Immunofluorescent staining 1:400 CST 13,110 Ki-67 (D3B5) 1:400 CST 9129 anti-rabbit IgG Alexa Fluor 488 1:200 Invitrogen A11008 anti-rabbit IgG DyLight TM 549 1:200 Vector laboratories DI-1549–1.5 P-AKT WB 1:500 CUSABIO CSB-PA008120 T-AKT 1:500 CUSABIO CSB-PA000855 P-ERK 1:1,000 CST 9101 s T-ERK 1:1,000 Assaybio B7074 T-FGFR1 1:1,000 Novus Biologicals NBP2-33784 P- FGFR1 (Tyr653, Tyr654) 1:1,000 Invitrogen 44-1140G T-FRS2 1:125 Biovision 3836–100 P-FRS2-alpha 1:1,000 CST 3864S IKKa 1:500 CST 3G12 b-actin 1:1,000 Santa cruz sc-8432 HRP linked anti-rabbit IgG 1:2,000 CST 7074 HRP linked anti-mouse IgG 1:2,000 CST 7076 FACS: Fluorescence-activated Cell Sorting. .. WB: Western blot Signaling Technology) and rabbit anti-Ki-67 (Cat No. 9129, Cell Signaling Technology), and the secondary antibody was DyLightTM 549 Goat Anti-Rabbit IgG (Cat No. DI-1549–1.5, Vector laboratories, Burlingame, CA, USA) and Alexa Fluor 488 goat anti-rabbit IgG (Cat No. A11008, Invitrogen) which were diluted in 3 % blocking buffer.

Western Blot:

Article Title: Efficient improvement of the proliferation, differentiation, and anti-arthritic capacity of mesenchymal stem cells by simply culturing on the immobilized FGF2 derived peptide, 44-ERGVVSIKGV-53.
Article Snippet: .. Antibody Assay Dilution Company source Catalog Number CD34 FACS 1:200 R&D system MAB72271 CD45 PD7/26/16 + 2B11- 1:200 Invitrogen MA5-13197 CD73/NT5E 1:200 Invitrogen RG235718 CD90/Thy1 1:200 R&D system AF2067 CD105 1:200 Invitrogen MA5-11854 anti-Mouse IgG Alexa Fluor 488 1:200 Invitrogen A11059 anti-Sheep lgG PE 1:200 R&D system F0126 PCNA (D3H8P) XP Immunofluorescent staining 1:400 CST 13,110 Ki-67 (D3B5) 1:400 CST 9129 anti-rabbit IgG Alexa Fluor 488 1:200 Invitrogen A11008 anti-rabbit IgG DyLight TM 549 1:200 Vector laboratories DI-1549–1.5 P-AKT WB 1:500 CUSABIO CSB-PA008120 T-AKT 1:500 CUSABIO CSB-PA000855 P-ERK 1:1,000 CST 9101 s T-ERK 1:1,000 Assaybio B7074 T-FGFR1 1:1,000 Novus Biologicals NBP2-33784 P- FGFR1 (Tyr653, Tyr654) 1:1,000 Invitrogen 44-1140G T-FRS2 1:125 Biovision 3836–100 P-FRS2-alpha 1:1,000 CST 3864S IKKa 1:500 CST 3G12 b-actin 1:1,000 Santa cruz sc-8432 HRP linked anti-rabbit IgG 1:2,000 CST 7074 HRP linked anti-mouse IgG 1:2,000 CST 7076 FACS: Fluorescence-activated Cell Sorting. .. WB: Western blot Signaling Technology) and rabbit anti-Ki-67 (Cat No. 9129, Cell Signaling Technology), and the secondary antibody was DyLightTM 549 Goat Anti-Rabbit IgG (Cat No. DI-1549–1.5, Vector laboratories, Burlingame, CA, USA) and Alexa Fluor 488 goat anti-rabbit IgG (Cat No. A11008, Invitrogen) which were diluted in 3 % blocking buffer.

Fluorescence:

Article Title: Efficient improvement of the proliferation, differentiation, and anti-arthritic capacity of mesenchymal stem cells by simply culturing on the immobilized FGF2 derived peptide, 44-ERGVVSIKGV-53.
Article Snippet: .. Antibody Assay Dilution Company source Catalog Number CD34 FACS 1:200 R&D system MAB72271 CD45 PD7/26/16 + 2B11- 1:200 Invitrogen MA5-13197 CD73/NT5E 1:200 Invitrogen RG235718 CD90/Thy1 1:200 R&D system AF2067 CD105 1:200 Invitrogen MA5-11854 anti-Mouse IgG Alexa Fluor 488 1:200 Invitrogen A11059 anti-Sheep lgG PE 1:200 R&D system F0126 PCNA (D3H8P) XP Immunofluorescent staining 1:400 CST 13,110 Ki-67 (D3B5) 1:400 CST 9129 anti-rabbit IgG Alexa Fluor 488 1:200 Invitrogen A11008 anti-rabbit IgG DyLight TM 549 1:200 Vector laboratories DI-1549–1.5 P-AKT WB 1:500 CUSABIO CSB-PA008120 T-AKT 1:500 CUSABIO CSB-PA000855 P-ERK 1:1,000 CST 9101 s T-ERK 1:1,000 Assaybio B7074 T-FGFR1 1:1,000 Novus Biologicals NBP2-33784 P- FGFR1 (Tyr653, Tyr654) 1:1,000 Invitrogen 44-1140G T-FRS2 1:125 Biovision 3836–100 P-FRS2-alpha 1:1,000 CST 3864S IKKa 1:500 CST 3G12 b-actin 1:1,000 Santa cruz sc-8432 HRP linked anti-rabbit IgG 1:2,000 CST 7074 HRP linked anti-mouse IgG 1:2,000 CST 7076 FACS: Fluorescence-activated Cell Sorting. .. WB: Western blot Signaling Technology) and rabbit anti-Ki-67 (Cat No. 9129, Cell Signaling Technology), and the secondary antibody was DyLightTM 549 Goat Anti-Rabbit IgG (Cat No. DI-1549–1.5, Vector laboratories, Burlingame, CA, USA) and Alexa Fluor 488 goat anti-rabbit IgG (Cat No. A11008, Invitrogen) which were diluted in 3 % blocking buffer.

RNA Sequencing:

Article Title: Pharmacological or genetic inhibition of LTCC promotes cardiomyocyte proliferation through inhibition of calcineurin activity
Article Snippet: .. LacZ RRAD 2F 2F/RRAD 0 200 400 600 800 RRAD Fo ld c ha ng e ✱✱ ns ns ✱✱ LacZ RRAD 2F 2F/RRAD 0 5000 10000 15000 20000 RRAD 2F CCND Fo ld c ha ng e ns ✱✱✱ ✱ ns LacZ RRAD 2F 2F/RRAD 0 1000 2000 3000 4000 5000 RRAD 2F CDK4 Fo ld c ha ng e ✱ ✱✱ ns ✱✱ a b c Supplementary Data 1: The RNA sequencing data was provided as an excel sheet in the supplementary material Supplementary Data 2: All the echocardiographic data from the MI experiment Supplementary Table 1: The antibodies used in the study for immunocytochemistry in hCOs Reagent Dilution/ concent ration Source Identifier Rabbit anti Ki-67 (D3B5) 1:400 Cell Signaling Technologies RRID:AB_2687446 Rabbit anti-phosphoHistone H3 (Ser10) 1:200 Merck Millipore RRID:AB_310177 Mouse anti-a-actinin, use 1:1000 1:1000 Sigma RRID:AB_476766 Mouse anti Ki-67 (8D5) 1:400 Cell Signaling Technologies RRID:AB_2797703 Rabbit anti-NKX 2.5 (E1Y8H) 1:400 Cell Signaling Technologies RRID:AB2797667 Goat anti-Mouse IgG (H+L) CrossAdsorbed Secondary Antibody, Alexa Fluor 488 1:400 ThermoFisher Scientific RRID: AB_2534069 Goat anti-Mouse IgG (H+L) CrossAdsorbed Secondary Antibody, Alexa Fluor 555 1:400 ThermoFisher Scientific RRID:AB_2535844 Goat anti-Rabbit IgG (H+L) CrossAdsorbed Secondary Antibody, Alexa Fluor 488, 1:400 ThermoFisher Scientific RRID:AB_143165 Goat anti-Rabbit IgG (H+L) CrossAdsorbed Secondary Antibody, Alexa Fluor 555 1:400 ThermoFisher Scientific RRID: AB_2535849 Hoechst33342 1:1000 Sigma - Supplementary Table 2: The antibodies used in the study for immunocytochemistry in NMCM P7, human heart slices, and fibroblast. ..

Immunocytochemistry:

Image Search Results

Effect of MR-39 on cell proliferation and protein expression in U87-MG cells. ( A ) Representative images of U87-MG cells treated for 48 h with either vehicle (CTRL) or 10 µM MR-39, followed by staining with DAPI and Ki67 antibody, to assess proliferation. ( B ) pERK protein levels, normalized to total ERK, and ( C ) pAKT protein levels, normalized to total AKT, were measured in U87MG cells both vehicle-treated (CTRL) and treated with 10 µM MR-39 for 48 h Western blot images show the molecular weight of each protein (in kDa) on the right. Protein expression is presented as a percentage of the CTRL. Scale bar, 25 μm. Data are expressed as mean ± SEM (n = 3). Statistical significance was determined by an unpaired t-test (**p < 0.01, ***p < 0.001)

Journal: Journal of Translational Medicine

Article Title: Formyl peptide receptor 2 activation by MR-39 inhibits glioblastoma cell proliferation and invasiveness through suppression of multiple oncogenic pathways

doi: 10.1186/s12967-026-07781-3

Figure Lengend Snippet: Effect of MR-39 on cell proliferation and protein expression in U87-MG cells. ( A ) Representative images of U87-MG cells treated for 48 h with either vehicle (CTRL) or 10 µM MR-39, followed by staining with DAPI and Ki67 antibody, to assess proliferation. ( B ) pERK protein levels, normalized to total ERK, and ( C ) pAKT protein levels, normalized to total AKT, were measured in U87MG cells both vehicle-treated (CTRL) and treated with 10 µM MR-39 for 48 h Western blot images show the molecular weight of each protein (in kDa) on the right. Protein expression is presented as a percentage of the CTRL. Scale bar, 25 μm. Data are expressed as mean ± SEM (n = 3). Statistical significance was determined by an unpaired t-test (**p < 0.01, ***p < 0.001)

Article Snippet: For Ki67 staining, cells were incubated overnight at 4 °C with Ki67 (D3B5) antibody rabbit mAb Alexa Fluor 488 Conjugate (1:500, Cell Signaling, Milan, Italy #11882).

Techniques: Expressing, Staining, Western Blot, Molecular Weight

FOXO4-DRI can effectively maintain vascular function in naturally aging mice. (A) Western blot experiments were conducted and quantitatively analyzed to determine the changes in protein levels of Ki-67, Lamin B, P21, P16, and γ-H2AX in the aortas of naturally aged mice after FOXO4-DRI or PBS treatment. n ≥ 4 per group. (B) RT-qPCR analysis was performed to examine the mRNA levels of Il-1β, Il-6, Cxcl15 , and Tnf-α in the aortas of naturally aged mice after FOXO4-DRI and PBS treatments. n = 3 per group. (C) Representative SA-β-Gal staining and quantitative analysis were performed to determine the number of positive cells in the aortas of naturally aged mice after FOXO4-DRI and PBS treatments. n = 5 per group. Scale bar = 100 μm. (D) Representative HE staining and quantitative analysis were performed to evaluate the thickness of the aortas in naturally aged mice after FOXO4-DRI and PBS treatments. n = 5 per group. Scale bar = 100 μm. (E) Color Doppler imaging and analysis were performed to detect and analyze the structural and blood flow conditions of the aortas in naturally aged mice after FOXO4-DRI and PBS treatments. n = 6 per group. (F) Representative DHE staining and quantitative analysis were performed to assess the ROS levels in the aortas of naturally aged mice after FOXO4-DRI and PBS treatments. n = 5 per group. Scale bar = 100 μm. Data are presented as mean ± SEM and analyzed using a two-tailed Student’s t-test. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001.

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: FOXO4-DRI regulates endothelial cell senescence via the P53 signaling pathway

doi: 10.3389/fbioe.2025.1729166

Figure Lengend Snippet: FOXO4-DRI can effectively maintain vascular function in naturally aging mice. (A) Western blot experiments were conducted and quantitatively analyzed to determine the changes in protein levels of Ki-67, Lamin B, P21, P16, and γ-H2AX in the aortas of naturally aged mice after FOXO4-DRI or PBS treatment. n ≥ 4 per group. (B) RT-qPCR analysis was performed to examine the mRNA levels of Il-1β, Il-6, Cxcl15 , and Tnf-α in the aortas of naturally aged mice after FOXO4-DRI and PBS treatments. n = 3 per group. (C) Representative SA-β-Gal staining and quantitative analysis were performed to determine the number of positive cells in the aortas of naturally aged mice after FOXO4-DRI and PBS treatments. n = 5 per group. Scale bar = 100 μm. (D) Representative HE staining and quantitative analysis were performed to evaluate the thickness of the aortas in naturally aged mice after FOXO4-DRI and PBS treatments. n = 5 per group. Scale bar = 100 μm. (E) Color Doppler imaging and analysis were performed to detect and analyze the structural and blood flow conditions of the aortas in naturally aged mice after FOXO4-DRI and PBS treatments. n = 6 per group. (F) Representative DHE staining and quantitative analysis were performed to assess the ROS levels in the aortas of naturally aged mice after FOXO4-DRI and PBS treatments. n = 5 per group. Scale bar = 100 μm. Data are presented as mean ± SEM and analyzed using a two-tailed Student’s t-test. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001.

Article Snippet: The primary antibodies used are as follows: P53 (Proteintech, 60283-2-Ig), pSer46-P53 (CST, 2521), FOXO4 (Proteintech, 21535-1-AP), Ki-67 (D3B5) Rabbit mAb (Alexa Fluor® 647 Conjugate) (CST, 12075), P21 (CST, 14074), γ-H2AX (Abcam, ab2254).

Techniques: Western Blot, Quantitative RT-PCR, Staining, Imaging, Two Tailed Test

FOXO4-DRI can effectively maintain vascular function in progeroid mice. (A) Western blot experiments were conducted and quantitatively analyzed to determine the changes in protein levels of Ki-67, Lamin B, P21, P16, and γ-H2AX in the aortas of progeroid mice after FOXO4-DRI or PBS treatment. n ≥ 4 per group. (B) RT-qPCR analysis was performed to examine the mRNA levels of Il-1β , Il-6 , Cxcl15 , and Tnf-α in the aortas of progeroid mice after FOXO4-DRI and PBS treatments. n = 3 per group. (C) Representative SA-β-Gal staining and quantitative analysis were performed to determine the number of positive cells in the aortas of progeroid mice after FOXO4-DRI and PBS treatments. Scale bar = 100 μm. n = 5 per group. (D) Representative HE staining and quantitative analysis were performed to evaluate the thickness in the aortas of progeroid mice after FOXO4-DRI and PBS treatments. Scale bar = 100 μm. n = 5 per group. (E) Color Doppler imaging and analysis were performed to detect and analyze the structural and blood flow conditions in the aortas of progeroid mice after FOXO4-DRI and PBS treatments. n = 5 per group. (F) Representative DHE staining and quantitative analysis were performed to assess the ROS levels in the aortas of progeroid mice after FOXO4-DRI and PBS treatments. n = 5 per group. Scale bar = 100 μm. Data are presented as mean ± SEM and analyzed using a two-tailed Student’s t-test. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001.

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: FOXO4-DRI regulates endothelial cell senescence via the P53 signaling pathway

doi: 10.3389/fbioe.2025.1729166

Figure Lengend Snippet: FOXO4-DRI can effectively maintain vascular function in progeroid mice. (A) Western blot experiments were conducted and quantitatively analyzed to determine the changes in protein levels of Ki-67, Lamin B, P21, P16, and γ-H2AX in the aortas of progeroid mice after FOXO4-DRI or PBS treatment. n ≥ 4 per group. (B) RT-qPCR analysis was performed to examine the mRNA levels of Il-1β , Il-6 , Cxcl15 , and Tnf-α in the aortas of progeroid mice after FOXO4-DRI and PBS treatments. n = 3 per group. (C) Representative SA-β-Gal staining and quantitative analysis were performed to determine the number of positive cells in the aortas of progeroid mice after FOXO4-DRI and PBS treatments. Scale bar = 100 μm. n = 5 per group. (D) Representative HE staining and quantitative analysis were performed to evaluate the thickness in the aortas of progeroid mice after FOXO4-DRI and PBS treatments. Scale bar = 100 μm. n = 5 per group. (E) Color Doppler imaging and analysis were performed to detect and analyze the structural and blood flow conditions in the aortas of progeroid mice after FOXO4-DRI and PBS treatments. n = 5 per group. (F) Representative DHE staining and quantitative analysis were performed to assess the ROS levels in the aortas of progeroid mice after FOXO4-DRI and PBS treatments. n = 5 per group. Scale bar = 100 μm. Data are presented as mean ± SEM and analyzed using a two-tailed Student’s t-test. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001.

Techniques: Western Blot, Quantitative RT-PCR, Staining, Imaging, Two Tailed Test

Glucose and oxygen deprivation can induce endothelial cell senescence. (A) Western blot analysis was performed to quantify the changes in protein levels of Ki-67, Lamin B, FOXO4, P53, P21, P16, and γ-H2AX in HUVECs following OGD treatment, with n ≥ 4 per group. (B) Representative immunofluorescence staining was conducted to analyze the content and localization changes of Ki-67, P21, γ-H2AX, and P53 different groups of HUVECs, with n = 5 per group. (C) Subcellular localization of FOXO4 in different groups of HUVECs was analyzed through representative immunofluorescence staining (FOXO4 in red, DAPI in blue), with n = 5 per group, scale bar = 40 μm. (D) Representative SA-β-GAL staining and quantification were performed to evaluate the number of positive cells in HUVECs after OGD treatment, with n = 5 per group, scale bar = 100 μm. (E) Tube formation assays analyzed relative tube lengths in different groups following OGD treatment, with n = 5 per group, scale bar = 400 μm. (F) Scratch assays were conducted to assess endothelial cell migration coverage at 0, 12, 24, and 36 h across different groups, with n = 5 per group, scale bar = 400 μm. Data are represented as mean ± SEM. Statistical analysis assessed by 2-way RM ANOVA (G) Representative DHE staining and quantification evaluated the number of DHE-positive cells in different groups, with n = 5 per group, scale bar = 100 μm. (H) RT-qPCR analysis was performed in HUVECs following OGD treatment to measure the mRNA levels of IL-6 , IL-8 , IL-1β , and TNF-α , with n = 3 per group. Data are presented as mean ± SEM and analyzed using a two-tailed Student’s t-test. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001.

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: FOXO4-DRI regulates endothelial cell senescence via the P53 signaling pathway

doi: 10.3389/fbioe.2025.1729166

Figure Lengend Snippet: Glucose and oxygen deprivation can induce endothelial cell senescence. (A) Western blot analysis was performed to quantify the changes in protein levels of Ki-67, Lamin B, FOXO4, P53, P21, P16, and γ-H2AX in HUVECs following OGD treatment, with n ≥ 4 per group. (B) Representative immunofluorescence staining was conducted to analyze the content and localization changes of Ki-67, P21, γ-H2AX, and P53 different groups of HUVECs, with n = 5 per group. (C) Subcellular localization of FOXO4 in different groups of HUVECs was analyzed through representative immunofluorescence staining (FOXO4 in red, DAPI in blue), with n = 5 per group, scale bar = 40 μm. (D) Representative SA-β-GAL staining and quantification were performed to evaluate the number of positive cells in HUVECs after OGD treatment, with n = 5 per group, scale bar = 100 μm. (E) Tube formation assays analyzed relative tube lengths in different groups following OGD treatment, with n = 5 per group, scale bar = 400 μm. (F) Scratch assays were conducted to assess endothelial cell migration coverage at 0, 12, 24, and 36 h across different groups, with n = 5 per group, scale bar = 400 μm. Data are represented as mean ± SEM. Statistical analysis assessed by 2-way RM ANOVA (G) Representative DHE staining and quantification evaluated the number of DHE-positive cells in different groups, with n = 5 per group, scale bar = 100 μm. (H) RT-qPCR analysis was performed in HUVECs following OGD treatment to measure the mRNA levels of IL-6 , IL-8 , IL-1β , and TNF-α , with n = 3 per group. Data are presented as mean ± SEM and analyzed using a two-tailed Student’s t-test. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001.

Techniques: Western Blot, Immunofluorescence, Staining, Migration, Quantitative RT-PCR, Two Tailed Test

FOXO4-DRI can alleviate endothelial cell senescence induced by OGD. (A) Western blot analysis was conducted to quantify the changes in protein levels of Ki-67, Lamin B, P21, P16, and γ-H2AX in senescent HUVECs following treatment with FOXO4-DRI or PBS, with n ≥ 4 per group. (B) Representative immunofluorescence analysis (green for Ki-67, blue for DAPI) was performed to assess the number of Ki-67 positive HUVECs in different groups, with n = 5 per group and scale bar = 40 μm. (C) Fluorescent staining was carried out to analyze the content and localization changes of P21 and γ-H2AX in different groups, with n = 5 per group and scale bar = 40 μm. (D) Representative SA-β-GAL staining and quantification were performed to evaluate the number of positive cells in different groups, with n = 5 per group and scale bar = 100 μm. (E) Tube formation assays were conducted to analyze relative tube lengths in different groups, with n = 5 per group and scale bar = 400 μm. (F) Representative DHE staining and quantification were carried out to evaluate the number of DHE-positive cells in different groups, with n = 5 per group and scale bar = 100 μm. (G) Scratch assays were performed to assess endothelial cell migration coverage at 0, 12, 24, and 36 h in different groups, with n = 5 per group and scale bar = 400 μm. Data are represented as mean ± SEM. Statistical analysis assessed by 2-way RM ANOVA. (H) RT-qPCR analysis was performed to detect mRNA levels of IL-6 , IL-8 , IL-1β , and TNF-α in different groups, with n = 3 per group. Data are presented as mean ± SEM and analyzed using a two-tailed Student’s t-test. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001.

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: FOXO4-DRI regulates endothelial cell senescence via the P53 signaling pathway

doi: 10.3389/fbioe.2025.1729166

Figure Lengend Snippet: FOXO4-DRI can alleviate endothelial cell senescence induced by OGD. (A) Western blot analysis was conducted to quantify the changes in protein levels of Ki-67, Lamin B, P21, P16, and γ-H2AX in senescent HUVECs following treatment with FOXO4-DRI or PBS, with n ≥ 4 per group. (B) Representative immunofluorescence analysis (green for Ki-67, blue for DAPI) was performed to assess the number of Ki-67 positive HUVECs in different groups, with n = 5 per group and scale bar = 40 μm. (C) Fluorescent staining was carried out to analyze the content and localization changes of P21 and γ-H2AX in different groups, with n = 5 per group and scale bar = 40 μm. (D) Representative SA-β-GAL staining and quantification were performed to evaluate the number of positive cells in different groups, with n = 5 per group and scale bar = 100 μm. (E) Tube formation assays were conducted to analyze relative tube lengths in different groups, with n = 5 per group and scale bar = 400 μm. (F) Representative DHE staining and quantification were carried out to evaluate the number of DHE-positive cells in different groups, with n = 5 per group and scale bar = 100 μm. (G) Scratch assays were performed to assess endothelial cell migration coverage at 0, 12, 24, and 36 h in different groups, with n = 5 per group and scale bar = 400 μm. Data are represented as mean ± SEM. Statistical analysis assessed by 2-way RM ANOVA. (H) RT-qPCR analysis was performed to detect mRNA levels of IL-6 , IL-8 , IL-1β , and TNF-α in different groups, with n = 3 per group. Data are presented as mean ± SEM and analyzed using a two-tailed Student’s t-test. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001.

Techniques: Western Blot, Immunofluorescence, Staining, Migration, Quantitative RT-PCR, Two Tailed Test

Review

ki 67 d3b5 (Cell Signaling Technology Inc)

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Incubation:

other:

Immunohistochemistry:

FACS:

Staining:

Western Blot:

Fluorescence:

RNA Sequencing:

Immunocytochemistry:

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