Journal: Nutrients

Article Title: The Effect of Resveratrol or Curcumin on Head and Neck Cancer Cells Sensitivity to the Cytotoxic Effects of Cisplatin

doi: 10.3390/nu12092596

Figure Lengend Snippet: The total p21 protein expression (pg/mL) in normal cell line-HUVEC and in tumor cell line-PE/CA-PJ49 cells treated 24 h with CisPt and/or RSV, CRM. The experiments were performed in triplicates. Results are expressed as mean values of three determinations ± standard deviation (SD). (** p < 0.005, *** p < 0.0005; **** p < 0.00005).

Article Snippet: DuoSetR IC ELISA-Human Total p21/CIP1/CDKN1A(: DYC1047-2) was purchased from R&D Systems Inc. and contains the basic components required for the development of sandwich ELISAs to measure human p21 protein also known as CIP1 and CDKN1A in cell lysates.

Techniques: Expressing, Standard Deviation

Journal: Nutrients

Article Title: The Effect of Resveratrol or Curcumin on Head and Neck Cancer Cells Sensitivity to the Cytotoxic Effects of Cisplatin

doi: 10.3390/nu12092596

Figure Lengend Snippet: The effects of CisPt, RSV, CRM treatment applied alone or in combination for 24 h on the level of p21 protein expression (n-fold p21protein expression) in normal cell line-Huvec and in tumor cell line-PE/CA-PJ49. The n-fold p21expression was calculated using the formula: n-fold p21expression = p21 pg/mL treatment/p21 pg/mL control.

Article Snippet: DuoSetR IC ELISA-Human Total p21/CIP1/CDKN1A(: DYC1047-2) was purchased from R&D Systems Inc. and contains the basic components required for the development of sandwich ELISAs to measure human p21 protein also known as CIP1 and CDKN1A in cell lysates.

Techniques: Expressing, Control

Journal: Nutrients

Article Title: The Effect of Resveratrol or Curcumin on Head and Neck Cancer Cells Sensitivity to the Cytotoxic Effects of Cisplatin

doi: 10.3390/nu12092596

Figure Lengend Snippet: The effect of treatment with CisPt, RSV, CRM applied independently or in combination on P21 gene expression in tumor cells PE/CA-PJ49 compared to normal cells HUVEC. Each sample was performed in duplicate. The samples were analyzed using the formula 2-ΔΔCt = gene expression. (* p < 0.05, ** p < 0.005; *** p < 0.0005).

Article Snippet: DuoSetR IC ELISA-Human Total p21/CIP1/CDKN1A(: DYC1047-2) was purchased from R&D Systems Inc. and contains the basic components required for the development of sandwich ELISAs to measure human p21 protein also known as CIP1 and CDKN1A in cell lysates.

Techniques: Gene Expression

Journal: Journal of Biological Chemistry

Article Title: Epidermal Growth Factor Receptor Pathway Analysis Identifies Amphiregulin as a Key Factor for Cisplatin Resistance of Human Breast Cancer Cells

doi: 10.1074/jbc.m706287200

Figure Lengend Snippet: FIGURE 3. Analysis of AKT kinase, downstream signaling reveals inactiva- tion of the p53 pathway in MCF-7 CisR cells. A, quantification of AKT kinase activity. To measure AKT kinase activity, a solid phase ELISA, which utilizes a specific synthetic peptide as a substrate and a polyclonal antibody that rec- ognizes the phosphorylated form of the substrate, was used. (n 3, ***, p 0.001). B, detection of p53 protein by immunoblotting (IB) using a polyclonal affinity-purified goat Ab specific for p53. MCF-7 cells (lane 1) and MCF-7 CisR cells (lane 2). Lane M, molecular weight marker. p53 is indicated by an arrow. C, quantification of p53 by a sandwich ELISA that measures human total p53 in cell lysates (n 3, ***, p 0.001). D, p21 expression indicates p53 pathway activity. Detection of p21 in whole cell lysates of MCF-7 (lane 1) and MCF-7 CisR cells (lane 2) by immunoblotting using a polyclonal affinity-purified goat Ab specific for p21. Lane M, molecular weight marker. p21 indicated by an arrow. E, to quantify the levels of p21 protein a sandwich ELISA that measures p21 in cell lysates was used. (n 3, ***, p 0.001). F, to quantify BCL-2 expression an ELISA that detects human BCL-2 in cell lysates was used (n 3, ***, p 0.001).

Article Snippet: For immunoblotting we used a polyclonal affinity-purified goat Ab specific for p53 at a concentration of 1 g/ml (AF1355, R & D Systems, Wiesbaden, Germany) and a polyclonal affinity-purified goat Ab specific for p21 at a concentration of 1 g/ml (AF1047, R & D Systems, Wiesbaden, Germany).

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Affinity Purification, Molecular Weight, Marker, Sandwich ELISA, Expressing

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 employed were: P53 (Proteintech, 60283-2-Ig), pSer46-P53 (CST, 2521), FOXO4 (Proteintech, 21535-1-AP), P21 (CST, 2947), P16 (CST, 18769), BAX (CST, 2772), BCL2 (CST, 3498), Ki-67 (Proteintech, 84192-4-RR), Lamin B (CST, 13435), Cleaved Caspase 3 (CST, 9664), γ-H2AX (Abcam, ab81299), and GAPDH (Abcam, ab8245) as an internal reference for normalizing protein expression levels.

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

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.

Article Snippet: The primary antibodies employed were: P53 (Proteintech, 60283-2-Ig), pSer46-P53 (CST, 2521), FOXO4 (Proteintech, 21535-1-AP), P21 (CST, 2947), P16 (CST, 18769), BAX (CST, 2772), BCL2 (CST, 3498), Ki-67 (Proteintech, 84192-4-RR), Lamin B (CST, 13435), Cleaved Caspase 3 (CST, 9664), γ-H2AX (Abcam, ab81299), and GAPDH (Abcam, ab8245) as an internal reference for normalizing protein expression levels.

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

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.

Article Snippet: The primary antibodies employed were: P53 (Proteintech, 60283-2-Ig), pSer46-P53 (CST, 2521), FOXO4 (Proteintech, 21535-1-AP), P21 (CST, 2947), P16 (CST, 18769), BAX (CST, 2772), BCL2 (CST, 3498), Ki-67 (Proteintech, 84192-4-RR), Lamin B (CST, 13435), Cleaved Caspase 3 (CST, 9664), γ-H2AX (Abcam, ab81299), and GAPDH (Abcam, ab8245) as an internal reference for normalizing protein expression levels.

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

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.

Article Snippet: The primary antibodies employed were: P53 (Proteintech, 60283-2-Ig), pSer46-P53 (CST, 2521), FOXO4 (Proteintech, 21535-1-AP), P21 (CST, 2947), P16 (CST, 18769), BAX (CST, 2772), BCL2 (CST, 3498), Ki-67 (Proteintech, 84192-4-RR), Lamin B (CST, 13435), Cleaved Caspase 3 (CST, 9664), γ-H2AX (Abcam, ab81299), and GAPDH (Abcam, ab8245) as an internal reference for normalizing protein expression levels.

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

Journal: International Journal of Molecular Sciences

Article Title: Lipopolysaccharide-Activated Macrophages Suppress Cellular Senescence and Promote Rejuvenation in Human Dermal Fibroblasts

doi: 10.3390/ijms26157061

Figure Lengend Snippet: Expression of the p16 and p21 genes in Young cells. The schematic protocol (1)–(7) is shown at the top of the figure. The test conditions in figures ( A – D ) correspond to the schematic protocol (1)–(7). Young cells were cultured for 24 h after replacing half of the medium with EMEM, Old cell-conditioned medium (Old CM), or THP/LPS-conditioned medium (THP/LPS-CM; 0–1000 ng/mL). ( A , B ) Total RNA was extracted, and the relative mRNA expression levels of p16 ( A ) and p21 ( B ) were measured by Quantitative real-time PCR (RT-qPCR) using GAPDH as a housekeeping gene. ( C , D ) Young cells were treated with EMEM or THP/LPS-CM (0, 10, or 100 ng/mL) for 24 h. Cells were lysed, and 12.5 μg of total protein per lane was separated by SDS-PAGE, followed by western blot analysis using antibodies against P16 ( C ) or P21 ( D ). Representative western blot images are shown in the boxed regions of panels C and D. Band intensities were normalized to β-actin and quantified. Data are presented as mean ± SD (n = 3). Different letters (a–c) indicate statistically significant differences ( p < 0.05, ANOVA, Tukey–Kramer).

Article Snippet: Proteins were resolved via 12.5% SDS-PAGE at 30 mA for 90 min and transferred onto PVDF membranes at 300 mA for 1 h. Membranes were then blocked with PBS containing 0.1% Tween-20 and 5% skim milk for 1 h, followed by incubation overnight at 4 °C with primary antibodies against P21 Waf1/Cip1 (1:1000) and β-actin (1:2000; both from Cell Signaling Technology, Danvers, MA, USA).

Techniques: Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, SDS Page, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Lipopolysaccharide-Activated Macrophages Suppress Cellular Senescence and Promote Rejuvenation in Human Dermal Fibroblasts

doi: 10.3390/ijms26157061

Figure Lengend Snippet: Changes in P16, P21, and Ki-67 expression in senescent NB1RGB cells treated with THP/LPS-CM. The schematic protocol (1)–(7) is shown at the top of the figure. The test conditions in figures ( A – D ) correspond to the schematic protocol (1)–(7). ( A ) Young and Old NB1RGB cells were treated with EMEM, THP/LPS (0 ng/mL)-CM, or THP/LPS (100 ng/mL)-CM for 24 h. Relative mRNA expression levels of p16 , p21 , and Ki-67 were analyzed by RT-qPCR and normalized to the levels in Young cells. ( B ) Old cells were treated with LPS (10, 100, or 1000 ng/mL) for 24 h, and the expression levels of p16 , p21 , and Ki-67 were assessed by RT-qPCR. In both ( A , B ), bars represent mean ± SD (n = 5). Different letters (a–c) indicate statistically significant differences ( p < 0.05). ( C ) Young and Old cells were treated with EMEM, THP/LPS (0 ng/mL)-CM, or THP/LPS (100 ng/mL)-CM for 24 h, then immunofluorescence staining was performed using antibodies against P21 and Ki-67 (n = 5). Representative fluorescence images were acquired using a 20× objective lens with 2× digital zoom. In P21 and Ki-67 single-staining panels green = P21 or Ki-67, red = actin, and blue = nuclei. In double-staining panels green = P21, red = Ki-67, and blue = nuclei. Scale bars = 50 μm. ( D ) Based on the immunofluorescence images, the percentages of P21-positive cells, Ki-67-positive cells, and total cell counts per field were quantified. Data are expressed as mean ± SD (n = 5). Different letters (a–c) indicate statistically significant differences ( p < 0.05, ANOVA, Tukey–Kramer).

Article Snippet: Proteins were resolved via 12.5% SDS-PAGE at 30 mA for 90 min and transferred onto PVDF membranes at 300 mA for 1 h. Membranes were then blocked with PBS containing 0.1% Tween-20 and 5% skim milk for 1 h, followed by incubation overnight at 4 °C with primary antibodies against P21 Waf1/Cip1 (1:1000) and β-actin (1:2000; both from Cell Signaling Technology, Danvers, MA, USA).

Techniques: Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, Fluorescence, Double Staining

Journal: Antioxidants

Article Title: Epicatechin Gallate Ameliorates UVB-Induced Photoaging by Inhibiting p38α-Mediated Autophagy and Oxidative Stress

doi: 10.3390/antiox15020180

Figure Lengend Snippet: Effects of ECG on UVB-induced senescence markers in HaCaT cells. ( A , B ) Cell cycle distribution analysis of HaCaT cells by flow cytometry. The histogram in ( A ) shows DNA content (propidium iodide staining intensity) on the x-axis and cell count on the y-axis. The stacked bar chart in ( B ) quantifies the percentage of cells in different cell cycle phases (G0/G1, S, and G2/M). ( C ) Relative mRNA expression levels of p53 , p16 , p21 , IL6 , and TNF-α determined by RT-qPCR. ( D , E ) Representative Western blots and densitometric analysis of p53, p16, and p21 proteins. ( F , I ) Representative immunofluorescence images of γH2AX and Lamin B1 (scale bar: 50 μm). γH2AX foci are shown in green, Lamin B1 is shown in red, and nuclei are counterstained with DAPI (blue). White arrows indicate nuclei with damage. ( G ) Representative images of SA-β-gal staining in HaCaT cells (scale bar: 200 μm). ( H ) Quantitative analysis of SA-β-gal positive cells. Data are presented as the mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001.

Article Snippet: Rabbit anti-Beclin1 (BECN1) antibody (cat. #11306), mouse anti-tubulin antibody (cat. #66031-1-IG), rabbit anti-LaminB1 (LMNB1) antibody (cat. #12987-1-AP), rabbit anti-Cyclin-Dependent Kinase Inhibitor 1A (p21, CDKN1A) antibody (cat. #10355-1-AP) and rabbit anti-LC3 antibody (cat. #14600) were purchased from Proteintech group (Wuhan, China).

Techniques: Flow Cytometry, Staining, Cell Characterization, Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence

Journal: Nature Communications

Article Title: Phosphorylation of PA28γ by CK2 kinase facilitates HNSCC tumor formation and progression

doi: 10.1038/s41467-025-67131-7

Figure Lengend Snippet: a IB analysis of WCLs derived from HEK293T cells transfected with His-E4F1 and increasing doses of Flag-PA28γ. b Quantitative real-time PCR analysis of the indicated genes in HEK293T cells transfected with EV and increasing doses of Flag-PA28γ. Endo., endogenous. c HSC-3 cells stably expressing EV or Flag-PA28γ were treated with 100 μgml −1 CHX for the indicated time before harvesting. Quantification of E4F1 levels relative to tubulin levels is shown. d , e HEK293T cells stably expressing EV or Flag-PA28γ were transfected with Myc-E4F1 truncations, and treated with 100 μgml −1 CHX for the indicated time before harvesting. Quantification of Myc-E4F1 levels relative to tubulin levels is shown. f HEK293T cells transfected with the indicated plasmids were treated without or with 10 μM MG132 for 6 h before harvesting, and WCLs were collected for IB analysis. g HEK293T cells stably expressing shRNA against endogenous PA28γ were transfected with the indicated plasmids, and WCLs were collected for IB analysis. h Purified PA28γ, E4F1 and 20S proteasome in the absence or presence of 100 nM proteasome inhibitor epoxomicin (Epox) were incubated as indicated for 45 min, followed by IB analysis. i Purified PA28γ WT, PA28γ-T23A, E4F1, p21 (as a positive control for E4F1) and 20S proteasome were incubated as indicated for 45 min, followed by IB analysis. Data in ( b – e ) represent the mean ± SD of three biological replicates; statistical significance was assessed by two-sided unpaired t -test. Source data are provided as a Source Data file.

Article Snippet: Recombinant human E4F1 protein (H00001877-P01) and recombinant human p21/CDKN1A protein (NBP2-22976) were purchased from Novus Biologicals.

Techniques: Derivative Assay, Transfection, Real-time Polymerase Chain Reaction, Stable Transfection, Expressing, shRNA, Purification, Incubation, Positive Control

Journal: Inflammation Research

Article Title: NLRP1 inflammasome promotes senescence and senescence-associated secretory phenotype

doi: 10.1007/s00011-024-01892-7

Figure Lengend Snippet: NLRP1 expression is associated with senescence. A Human fibroblasts were exposed to 20 Gy ionizing irradiation (IR). On day 1, 5 and 7, NLRP1 and senescence protein expression were analyzed by immunoblotting. B Representative images of Ki67 and NLRP1 immunofluorescence. Scale bar = 50 μm. C , D IL-1β, IL18, IL-6 and IL-8 were quantified by ELISA. All data are presented as means ± SEM, n = 4 independent experiments; ** P < 0.005, *** P < 0.001 differences between time points after irradiation and day 0. E , F Human fibroblasts were treated with Valboropro (Vbpro) to induce NLRP1 expression. After 24 h, NLRP1 and IL-6 protein expression were analyzed by immunoblotting and cytokines were analyzed by ELISA. Human fibroblasts were irradiated ( G ) or stimulated with palbociclib (Palbo) ( H ) to induce two different senescence models. Then, cells were transfected with a non-targeting control siRNA (Control) or with siRNAs against NLRP1 (siNLRP1). Expression of NLRP1 and senescence-associated proteins p16, p21, p53 and IL-6 were assessed by immunoblotting and IL6, IL-8 or IL-18 were quantified by ELISA. All data are presented as means ± SEM, n = 4 independent experiments; ** P < 0.005, *** P < 0.001 irradiated vs. control. aaa P < 0.001, IR + siRNA vs. IR cells (color figure online)

Article Snippet: Monoclonal antibodies specific for NLRP1 (NBP1-54899), NLRP3 (NBP2-12446) and p21/CIP1/CDKN1A (NBP2-29463) were purchased from Novus Biologicals (Colorado, USA).

Techniques: Expressing, Irradiation, Western Blot, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Transfection, Control

Journal: Inflammation Research

Article Title: NLRP1 inflammasome promotes senescence and senescence-associated secretory phenotype

doi: 10.1007/s00011-024-01892-7

Figure Lengend Snippet: NLRP1 contributes to cellular senescence in vivo. A Nlrp1 protein expression in liver from WT mice at 1 month after IR. B Serum levels of IL-18 after IR. C Effect of IR on the bodyweight of WT and Nlrp1 knockout (KO) mice. D Protein expression in liver from IR and non-IR WT and Nlrp1 KO mice of senescent markers (IL-6, p16 and p21). Densitometry in Supplementary Fig. 6. E Serum levels of IL-6 in serum from IR and non-IR WT and Nlrp1 KO mice. F Heat map depicting expression of 44 mouse cytokines in serum at 5 weeks after IR of WT and Nlrp1 KO mice. n = 6 mice per group. G IL-6 releases from healthy fibroblasts was assessed after 24 and 48 h of incubation with media containing serum from IR and non-IR WT and Nlrp1 KO mice. H Representative liver section stainings of hematoxylin and eosin (H&E). I Representative liver section immunostainings of NLRP1 and p16. All data are presented as means ± SEM, n = 6–8 mice per group; * P < 0.05, ** P < 0.005, *** P < 0.001 irradiated vs. control. aa P < 0.005 IR WT vs. IR KO mice; bb P < 0.005 IR KO vs. IR KO mice (color figure online)

Article Snippet: Monoclonal antibodies specific for NLRP1 (NBP1-54899), NLRP3 (NBP2-12446) and p21/CIP1/CDKN1A (NBP2-29463) were purchased from Novus Biologicals (Colorado, USA).

Techniques: In Vivo, Expressing, Knock-Out, Incubation, Irradiation, Control

Journal: Inflammation Research

Article Title: NLRP1 inflammasome promotes senescence and senescence-associated secretory phenotype

doi: 10.1007/s00011-024-01892-7

Figure Lengend Snippet: NLRP1 senses DNA damage dependent of cGAS activation. A Protein expression levels of NLRP1 and cGAS after 24 h exposition to gDNA from non-irradiated and irradiated cells. B IL-6 and IL-18 release to the medium after the same experimental condition. Levels were determined by ELISA assay. C , D Protein expression levels of NLRP1 and cGAS and IL-6 and IL-18 release after 24 h exposition to a non-irradiated or irradiated synthetic double-stranded DNA sequence, poly(dA-dT), Cytokine levels were determined by ELISA assay. Cytokine levels were determined by ELISA assay. All data are presented as means ± SEM, n = 4 independent experiments; ** P < 0.005, *** P < 0.001 gDNA vs. control cells. a P < 0.05, aa P < 0.005, aaa P < 0.001, IR gDNA vs. control cells. E Human fibroblasts were irradiated to induces senescence. Then, cells were transfected with a non-targeting control siRNA (Control) or with siRNAs against cGAS (sicGAS). Expression of NLRP1, NLRP3, IL-1β, cGAS and senescent protein p16, p21 was assessed by immunoblotting. F IL-18 release of non-irradiated, irradiated and irradiated and transfected with siRNAs against cGAS. Cytokine levels were determined by ELISA assay. All data are presented as means ± SEM, n = 4 independent experiments; ** P < 0.005, IR vs. control cells. aa P < 0.005, IR vs. sicGAS (color figure online)

Article Snippet: Monoclonal antibodies specific for NLRP1 (NBP1-54899), NLRP3 (NBP2-12446) and p21/CIP1/CDKN1A (NBP2-29463) were purchased from Novus Biologicals (Colorado, USA).

Techniques: Activation Assay, Expressing, Irradiation, Enzyme-linked Immunosorbent Assay, Sequencing, Control, Transfection, Western Blot