rabbit anti p53 primary antibody  (Proteintech)

Journal: iScience

Article Title: MDM2/MDMX inhibition by Sulanemadlin synergizes with anti-Programmed Death 1 immunotherapy in wild-type p53 tumors

doi: 10.1016/j.isci.2024.109862

Figure Lengend Snippet: Stapled peptide Sulanemadlin and small molecule MDM2 inhibitors Siremadlin and Navtemadlin efficiently inhibit cell growth p53 dependently in vitro (A–F) CT26.WT murine colon carcinoma cells, B16-F10 p53 +/+ and B16-F10 p53 −/− murine malignant melanoma cells were treated with indicated concentrations of Sulanemadlin and Siremadlin and cell growth was monitored in the Incucyte S3 live cell imaging system. Proliferation was calculated as % of confluence over time. IC 50 values were normalized to DMSO control and calculated after 96 h treatment. Representative data from at least two experiments performed in triplicates and a mean IC50 from all performed experiments are shown. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. One-way ANOVA, (data are represented as mean with SD). See also Figures S1 and .

Article Snippet: Anti-p53 monoclonal rabbit anti-mouse (primary) , Abcam , Cat#ab246223.

Techniques: In Vitro, Live Cell Imaging

Journal: iScience

Article Title: MDM2/MDMX inhibition by Sulanemadlin synergizes with anti-Programmed Death 1 immunotherapy in wild-type p53 tumors

doi: 10.1016/j.isci.2024.109862

Figure Lengend Snippet: Activation of p53 by inhibition of MDM2/MDMX with Sulanemadlin leads to increased expression of immunogenicity markers CT26.WT cells were treated with 4 μmol/L Sulanemadlin for 48 h and flow cytometry analysis of protein expression determined the median fluorescence intensity (MFI) of (A) p53, (B) p21, (C) PUMA, (D) MHC I, (E) RAE-1, (F) PD-L1. Gating according to FMO controls excluded debris and only included single live cells. Representative data from two separate experiments performed in triplicates. ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. One-way ANOVA, (data are represented as mean with SD). See also Figures S3 and .

Article Snippet: Anti-p53 monoclonal rabbit anti-mouse (primary) , Abcam , Cat#ab246223.

Techniques: Activation Assay, Inhibition, Expressing, Flow Cytometry, Fluorescence

Journal: iScience

Article Title: MDM2/MDMX inhibition by Sulanemadlin synergizes with anti-Programmed Death 1 immunotherapy in wild-type p53 tumors

doi: 10.1016/j.isci.2024.109862

Figure Lengend Snippet:

Article Snippet: Anti-p53 monoclonal rabbit anti-mouse (primary) , Abcam , Cat#ab246223.

Techniques: Recombinant

Journal: International Journal of Molecular Sciences

Article Title: The Role of Hydrogen Sulfide in the Localization and Expression of p53 and Cell Death in the Nervous Tissue in Traumatic Brain Injury and Axotomy

doi: 10.3390/ijms242115708

Figure Lengend Snippet: Immunofluorescence microscopy: ( a ) expression of p53 (green fluorescence) in the cerebral cortex of the control group, which was injected with saline, and experimental groups of animals that were injected with Na 2 S and AOAA, 24 h and 7 days after TBI. Scale bar 20 µm. ( b ) Expression of p53 (green fluorescence) in the cerebral cortex of the control group, which was injected with saline, and experimental groups of animals that were injected with Na 2 S and AOAA, 7 days after TBI. Scale bar 25 µm. ( c ) p53 expression (green fluorescence) in a brain section from an animal injected with AOAA, 7 days after TBI. The white square is the area for analysis of p53 expression. White arrow is the location of the injury. Scale bar 500 µm. ( d ) Coefficient M1 of colocalization of p53 and neuronal nuclear marker NeuN in the contralateral and ipsilateral cortex of the control and experimental groups 24 h and 7 days after TBI. ( e ) Dependence of the average intensity of p53 fluorescence in neurons of the contralateral and ipsilateral cortex of the control and experimental groups 24 h and 7 days after TBI. Contra, contralateral cortex; Ipsi, ipsilateral cortex. NeuN, marker of neuron nuclei (red fluorescence); NeuN+p53 and Hoechst+p53, overlap. Hoechst—Hoechst 33342 fluorescence (blue fluorescence), which visualizes the nuclei of all cells, neurons and glia. One-way ANOVA. M ± SEM. n = 6. ** p < 0.01, *** p < 0.001—ipsilateral cortex relative to contralateral cortex of one animal; # p < 0.05, ## p < 0.01—ipsilateral cortex of the experimental group versus ipsilateral cortex of the control group within one time period after injury.

Article Snippet: Sections are then incubated with rabbit primary anti-p53 antibody (1:100; PAA928Mu01, Cloud-Clone Corp, Wuhan, China) and mouse anti-neuN protein antibody (1:1000; FNab10266, FineTest, Wuhan, China) or astrocyte marker GFAP (1:1000; SAB4200571, Sigma Aldrich) for two days at 4 °C.

Techniques: Immunofluorescence, Microscopy, Expressing, Fluorescence, Injection, Saline, Marker

Journal: International Journal of Molecular Sciences

Article Title: The Role of Hydrogen Sulfide in the Localization and Expression of p53 and Cell Death in the Nervous Tissue in Traumatic Brain Injury and Axotomy

doi: 10.3390/ijms242115708

Figure Lengend Snippet: Immunofluorescence microscopy: ( a ) expression of p53 (green fluorescence) in the cerebral cortex of the control group, which was injected with saline, and experimental groups of animals that were injected with Na 2 S and AOAA, 24 h and 7 days after TBI. Scale bar 20 µm. ( b ) Expression of p53 (green fluorescence) in the cerebral cortex of the control group, which was injected with saline, and experimental groups of animals that were injected with Na 2 S and AOAA, 7 days after TBI. Scale bar 25 µm. ( c ) p53 expression (green fluorescence) in a brain section from an animal injected with AOAA, 7 days after TBI. The white square is the area for analysis of p53 expression. White arrow is the location of the injury. Scale bar 500 µm. ( d ) Coefficient M1 of p53 colocalization and GFAP neuronal nuclear marker in the contralateral and ipsilateral cortex of the control and experimental groups 24 h and 7 days after TBI. Scale bar 20 µm. Contra, contralateral cortex; Ipsi, ipsilateral cortex. GFAP+p53 and Hoechst+p53—overlap. Hoechst—Hoechst 33342 (blue fluorescence) fluorescence, which visualizes the nuclei of all cells, neurons and glia. One-way ANOVA. M ± SEM. n = 6. * p < 0.05, ** p < 0.01, *** p < 0.001—ipsilateral cortex relative to contralateral cortex of one animal; # p < 0.05, ## p < 0.01—ipsilateral cortex of the experimental group versus ipsilateral cortex of the control group within the same time period after injury.

Article Snippet: Sections are then incubated with rabbit primary anti-p53 antibody (1:100; PAA928Mu01, Cloud-Clone Corp, Wuhan, China) and mouse anti-neuN protein antibody (1:1000; FNab10266, FineTest, Wuhan, China) or astrocyte marker GFAP (1:1000; SAB4200571, Sigma Aldrich) for two days at 4 °C.

Techniques: Immunofluorescence, Microscopy, Expressing, Fluorescence, Injection, Saline, Marker

Journal: International Journal of Molecular Sciences

Article Title: The Role of Hydrogen Sulfide in the Localization and Expression of p53 and Cell Death in the Nervous Tissue in Traumatic Brain Injury and Axotomy

doi: 10.3390/ijms242115708

Figure Lengend Snippet: Western blot analysis. Effect of Na 2 S and AOAA on p53 protein expression in the contralateral and ipsilateral cortex 24 h and 7 days after TBI. One-way ANOVA. M ± SEM. n = 6. * p < 0.05, ** p < 0.01—ipsilateral cortex relative to contralateral cortex of one animal; # p < 0.05, ## p < 0.01—ipsilateral cortex of the experimental group versus ipsilateral cortex of the control group within one time period after injury.

Article Snippet: Sections are then incubated with rabbit primary anti-p53 antibody (1:100; PAA928Mu01, Cloud-Clone Corp, Wuhan, China) and mouse anti-neuN protein antibody (1:1000; FNab10266, FineTest, Wuhan, China) or astrocyte marker GFAP (1:1000; SAB4200571, Sigma Aldrich) for two days at 4 °C.

Techniques: Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: The Role of Hydrogen Sulfide in the Localization and Expression of p53 and Cell Death in the Nervous Tissue in Traumatic Brain Injury and Axotomy

doi: 10.3390/ijms242115708

Figure Lengend Snippet: Fluorescence microscopy. ( a ) Crayfish stretch receptor 6 h after axotomy, stained with Hoechst and propidium iodide, control and experimental groups, which were incubated with Na 2 S and AOAA. White dotted square—MRN; N—neuron nucleus; the white arrow—glial cells. ( b ) MRN axon section 6 h after axotomy, stained with Hoechst and propidium iodide of the control and experimental groups. White star—glial cell apoptosis. ( c ) Necrosis of glial cells surrounding the neuron body, 6 h after axotomy. ( d ) Apoptosis of glial cells MRN in a 2 mm area along the axon, was assessed by the number of fragmented glial cells. Scale bar 40 µm. Hoechst+p53—overlap. Hoechst—Hoechst 33342 (blue fluorescence) fluorescence, which visualizes the nuclei of all cells, neurons and glia. One-way ANOVA. M ± SEM. n = 6. * p < 0.05.

Article Snippet: Sections are then incubated with rabbit primary anti-p53 antibody (1:100; PAA928Mu01, Cloud-Clone Corp, Wuhan, China) and mouse anti-neuN protein antibody (1:1000; FNab10266, FineTest, Wuhan, China) or astrocyte marker GFAP (1:1000; SAB4200571, Sigma Aldrich) for two days at 4 °C.

Techniques: Fluorescence, Microscopy, Staining, Incubation

Journal: International Journal of Molecular Sciences

Article Title: The Role of Hydrogen Sulfide in the Localization and Expression of p53 and Cell Death in the Nervous Tissue in Traumatic Brain Injury and Axotomy

doi: 10.3390/ijms242115708

Figure Lengend Snippet: Immunofluorescence microscopy. ( a ) Expression of p53 (green fluorescence) in the stretch receptor of crayfish of the control group and experimental groups that were incubated with Na 2 S and AOAA, 6 after axotomy. ( b ) p53 expression (green fluorescence) in the distal axon region of the MRN of the control group and experimental groups that were incubated with Na 2 S and AOAA, 6 after axotomy. ( c ) p53 fluorescence intensity in the nucleus and cytoplasm of MRN, as well as in the nucleoplasm of glial cells 6 h after axotomy. ( d ) p53 fluorescence intensity in the proximal and distal portion of the axon, as well as in the dendritic region of the MRN. ( e ) Expression of p53 in the stretch receptor of crayfish and distal portion of the axon. ( f ) Crayfish stretch receptor in transmitted light. ( g ) Schematic of the crayfish stretch receptor. RM—receptor muscle; N—neuron nucleus; C—cytoplasm; D—dendrites; A—axon; G and white stars—glial cells; red arrows—p53 cord inside the axon; Hoechst+p53—overlap. Hoechst-Hoechst 33342 fluorescence, which visualizes the nuclei of all cells, neurons and glia. Scale bar 40 µm. One-way ANOVA. M ± SEM. n = 6. * p < 0.05, ** p < 0.01, *** p < 0.01.

Article Snippet: Sections are then incubated with rabbit primary anti-p53 antibody (1:100; PAA928Mu01, Cloud-Clone Corp, Wuhan, China) and mouse anti-neuN protein antibody (1:1000; FNab10266, FineTest, Wuhan, China) or astrocyte marker GFAP (1:1000; SAB4200571, Sigma Aldrich) for two days at 4 °C.

Techniques: Immunofluorescence, Microscopy, Expressing, Fluorescence, Incubation

Journal: International Journal of Molecular Sciences

Article Title: The Role of Hydrogen Sulfide in the Localization and Expression of p53 and Cell Death in the Nervous Tissue in Traumatic Brain Injury and Axotomy

doi: 10.3390/ijms242115708

Figure Lengend Snippet: Western blot analysis. Effect of Na 2 S and AOAA on p53 protein expression in axotomized ganglia of the ventral nerve cord 6 h after axotomy. One-way ANOVA. M ± SEM. n = 6. * p < 0.05, ** p < 0.01.

Article Snippet: Sections are then incubated with rabbit primary anti-p53 antibody (1:100; PAA928Mu01, Cloud-Clone Corp, Wuhan, China) and mouse anti-neuN protein antibody (1:1000; FNab10266, FineTest, Wuhan, China) or astrocyte marker GFAP (1:1000; SAB4200571, Sigma Aldrich) for two days at 4 °C.

Techniques: Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: The Role of Hydrogen Sulfide in the Localization and Expression of p53 and Cell Death in the Nervous Tissue in Traumatic Brain Injury and Axotomy

doi: 10.3390/ijms242115708

Figure Lengend Snippet: Conceptual scheme of the role of H 2 S in the regulation of p53 levels in TBI and axotomy. Arrows with a sharp end—positive regulation; arrows with a blunt end—negative regulation.

Article Snippet: Sections are then incubated with rabbit primary anti-p53 antibody (1:100; PAA928Mu01, Cloud-Clone Corp, Wuhan, China) and mouse anti-neuN protein antibody (1:1000; FNab10266, FineTest, Wuhan, China) or astrocyte marker GFAP (1:1000; SAB4200571, Sigma Aldrich) for two days at 4 °C.

Techniques:

Journal: Frontiers in Aging

Article Title: Metformin mitigates SASP secretion and LPS-triggered hyper-inflammation in Doxorubicin-induced senescent endothelial cells

doi: 10.3389/fragi.2023.1170434

Figure Lengend Snippet: Metformin inhibited DOX-induced upregulation of senescence markers and SA-β-gal activity in endothelial cells. (A) Schematic diagram of the experimental design. Both EA.hy926 endothelial derived cells and HUVECs were treated for 24 h with 0.5 µM DOX ± metformin (0.5–5 mM, added 24 h before DOX). Thereafter, DOX was removed and the cells were incubated in DOX-free media with or without metformin for an additional 72 h for protein expression experiments or 120 h for measurement of SA-β-gal staining. Expression levels of senescence markers including p-p53, p53, and p21 in EA.hy926 cells (B–D), and HUVECs [ (E–G) , respectively] were measured using western blot (n = 4–8). Representative images of western blots are shown. Values were normalized to α-tubulin and expressed relative to cells treated with DOX alone. (H) Images of SA-β-gal staining in control, DOX-treated, and DOX + metformin co-treated cells are shown in HUVECs. Images were analyzed and the percentage of SA-β-gal positive cells were calculated (n = 6–8). Values are presented as means ± SEM. Data were analyzed by one-way ANOVA followed by a Dunnet’s multiple comparisons test or non-parametric Kruskal–Wallis tests followed by Dunn’s post hoc test ; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Schematic diagram created with BioRender.com .

Article Snippet: Primary mouse antibodies against p53 (catalog 2,524, 1:1000 dilution) and primary rabbit antibodies against phospho-p53 (Ser15) (catalog 9284, 1:1000 dilution), p21 (catalog 2,947, 1:1000 dilution), MMP-3 (catalog 14,351, 1:1000 dilution), ICAM-1 (catalog 4915, 1:1000 dilution), phospho-SAPK/JNK (Thr183/Tyr185) (catalog 4668, 1:1000 dilution), SAPK/JNK (catalog 9252, 1:1000 dilution), phospho-p38 (Thr180/Tyr182) (catalog 4511, 1:1000 dilution), p38 (catalog 8690, 1:1000 dilution), AMPK alpha (catalog 2,532, 1:1000 dilution), phospho-NF-κB p65 (catalog 3033, 1:1000 dilution), and alpha-tubulin (catalog 2144, 1:1000 dilution) were purchased from Cell Signaling Technology (Danvers, MA, United States).

Techniques: Activity Assay, Derivative Assay, Incubation, Expressing, Staining, Western Blot, Control

Journal: Aging Cell

Article Title: Antiretroviral protease inhibitors induce features of cellular senescence that are reversible upon drug removal

doi: 10.1111/acel.13750

Figure Lengend Snippet: The anti‐HIV PI cocktail atazanavir + ritonavir (ATV/r) induces senescence in cultured cells. IMR‐90 primary human fibroblasts were cultured in the presence of ATV/r for 14 days. (a). RNA was isolated from untreated (DMSO) and ATV/r‐treated cells, and p16 INK4a and p21 WAF1 mRNA levels, normalized for Actin mRNA, were measured by qPCR. (b). mRNA levels of LMNB1 in DMSO‐ and ATV/r‐treated cells were similarly measured. (c). SASP component mRNA levels were measured using qPCR. (d). Representative images of SA‐β‐gal positivity in control cells (left panel) and cells induced to senesce by ATV/r (right panel). (e). Intracellular levels of proteins prelamin a, activated (P‐ser37) p53, p21 WAF1 , HMGB1 and beta‐Actin (control) using western analysis. (f). Cells were analyzed for HMGB1 release from the nucleus, proliferation (EdU), and nuclei morphology (DAPI) by microscopy.

Article Snippet: Separated proteins were transferred to polyvinylidene fluoride membranes, which were blocked and incubated overnight at 4°C with anti‐rabbit primary antibodies (p21 WAF1 , Cell Signaling #2947, 1:1000; p‐p53, Cell Signaling #9289, 1:1000; HMGB1, abcam# ab18256, 1:2000; prelamin A, Millipore Sigma #mabt858, 1:500; β‐actin, Sigma‐Aldrich #A2228, 1:10000).

Techniques: Cell Culture, Isolation, Control, Western Blot, Microscopy

Journal: Aging Cell

Article Title: Antiretroviral protease inhibitors induce features of cellular senescence that are reversible upon drug removal

doi: 10.1111/acel.13750

Figure Lengend Snippet: The HIV PI darunavir does not induce senescence in culture. Mouse p16‐3MR dermal fibroblasts were treated with DMSO (control), ATV/r or DRN/r for 14 d prior to analyses. (a). mRNA levels of p16 INK4a and p21 WAF1 were measured by qPCR. (b). mRNA levels of LMNB1 in treated cells as per (a). (c). Representative images of SA‐β‐gal positivity in control cells (DMSO), and DRN/r‐ and ATV/r‐treated cells. (d). Quantification of SA‐β‐gal positivity. (e). SASP component mRNA levels were measured by qPCR in the indicated cell populations. (f). Cell proliferation of DMSO‐, DRN/r‐ and ATV/r‐treated cells. Cell proliferation was followed for 4 days using the CellTrace™ violet reagent.

Article Snippet: Separated proteins were transferred to polyvinylidene fluoride membranes, which were blocked and incubated overnight at 4°C with anti‐rabbit primary antibodies (p21 WAF1 , Cell Signaling #2947, 1:1000; p‐p53, Cell Signaling #9289, 1:1000; HMGB1, abcam# ab18256, 1:2000; prelamin A, Millipore Sigma #mabt858, 1:500; β‐actin, Sigma‐Aldrich #A2228, 1:10000).

Techniques: Control

Journal: Aging Cell

Article Title: Antiretroviral protease inhibitors induce features of cellular senescence that are reversible upon drug removal

doi: 10.1111/acel.13750

Figure Lengend Snippet: ATV/r‐induces a p53‐dependent growth arrest that is reversed upon drug removal. IMR‐90 fibroblasts were transduced with lentiviral vectors expressing GSE22, a peptide that inactivates p53, or no insert (vector), and cultured in the presence of ATV/r or DRN/r for 14 days. (a). p21 WAF1 and LMNB1 mRNA levels in control (vector) cells were measured by qPCR. (b). p21 WAF1 and LMNB1 mRNA levels in GSE‐transduced cells were measured by qPCR. (c). Representative images of SA‐β‐gal positivity in control (vector) or GSE22‐transduced cells treated with ATV/r. (d). Cell proliferation of DMSO‐, ATV/r‐, and DRN/r‐treated (Lenti‐GSE22‐transduced) IMR‐90 cells was analyzed. Cell proliferation was followed for 4 days using the CellTrace™ violet reagent. (e). Western blot confirming p53 inactivation in GSE22‐transduced cells by absence of p21 WAF1 expression and retained nuclear HMGB1. (f). IMR‐90 fibroblasts were treated with DMSO, ATV/r or DRN/r for 14 d prior to analysis. p16 INK4a , p21 WAF1 and LMNB1 mRNA levels in the three cell populations. (g). Representative images and quantification of SA‐β‐gal positivity in DMSO‐treated, ATV/r‐treated or ATV/r‐released cells. (h). Proliferation of DMSO‐, DRN/r‐ and ATV/r‐treated cells was followed for 4 days using the CellTrace™ violet reagent. (i). the 17.1% non‐dividing ATV/r‐treated cells were re‐plated and cultured for 2 weeks without ATV/r, then re‐analyzed for proliferation. (j). Percentage of individual cells from DMSO‐ or ATV/r‐treated cultures that had divided by 96 h. (k). Images of individually tracked DMSO‐treated, ATV/r‐treated or ATV/r‐released cells. Magnification and scale are consistent between treatment groups.

Article Snippet: Separated proteins were transferred to polyvinylidene fluoride membranes, which were blocked and incubated overnight at 4°C with anti‐rabbit primary antibodies (p21 WAF1 , Cell Signaling #2947, 1:1000; p‐p53, Cell Signaling #9289, 1:1000; HMGB1, abcam# ab18256, 1:2000; prelamin A, Millipore Sigma #mabt858, 1:500; β‐actin, Sigma‐Aldrich #A2228, 1:10000).

Techniques: Transduction, Expressing, Plasmid Preparation, Cell Culture, Control, Western Blot

Journal: Aging Cell

Article Title: Antiretroviral protease inhibitors induce features of cellular senescence that are reversible upon drug removal

doi: 10.1111/acel.13750

Figure Lengend Snippet: ATV/r treatment accelerates aging phenotypes. Young adult mice at 5 months of age were treated with 62 mg/kg atazanavir and 21 mg/kg ritonavir in drinking water for 8 weeks. (a). Schematic of the experimental setup. (b). Representative phenotypic differences between ATV/r‐treated mice and age‐matched controls are shown. (c–h). p21 Waf1 mRNA levels were measured by qPCR in various tissues. (i). Representative images of p16‐3MR male mice injected with coelenterazine and measured for luminescence using the Xenogen imaging system. (j). Quantification of luminescence presented in I.

Article Snippet: Separated proteins were transferred to polyvinylidene fluoride membranes, which were blocked and incubated overnight at 4°C with anti‐rabbit primary antibodies (p21 WAF1 , Cell Signaling #2947, 1:1000; p‐p53, Cell Signaling #9289, 1:1000; HMGB1, abcam# ab18256, 1:2000; prelamin A, Millipore Sigma #mabt858, 1:500; β‐actin, Sigma‐Aldrich #A2228, 1:10000).

Techniques: Injection, Imaging

Journal: Aging Cell

Article Title: Antiretroviral protease inhibitors induce features of cellular senescence that are reversible upon drug removal

doi: 10.1111/acel.13750

Figure Lengend Snippet: ATV/r‐treated mice accumulate senescent cells at sites of age‐related pathologies. Young adult mice (5 months of age) were treated with 62 mg/kg atazanavir and 21 mg/kg ritonavir in drinking water for 8 weeks. (a). Representative images of SA‐β‐gal staining in subcutaneous fat from vehicle and ATV/r‐treated mice. (b). Quantification of SA‐β‐gal staining presented in a. (c–f). selected SASP factors (MMP3, IL‐1a, GMCSF and IL‐10) and p21 Waf1 mRNA expression levels were measured in subcutaneous fat by qPCR and normalized to Actin. (g). Representative images of sectioned dorsal skin stained for SA‐β‐gal activity. (h). qPCR analysis for senescence marker p16 Ink4a . (i–k). mRNA expression levels of some SASP markers from skin (IL‐10, TIMP1 and MMP3) were measured by qPCR and normalized to Actin. (l–n). cardiac activity was measured by echocardiography.

Article Snippet: Separated proteins were transferred to polyvinylidene fluoride membranes, which were blocked and incubated overnight at 4°C with anti‐rabbit primary antibodies (p21 WAF1 , Cell Signaling #2947, 1:1000; p‐p53, Cell Signaling #9289, 1:1000; HMGB1, abcam# ab18256, 1:2000; prelamin A, Millipore Sigma #mabt858, 1:500; β‐actin, Sigma‐Aldrich #A2228, 1:10000).

Techniques: Staining, Expressing, Activity Assay, Marker

Journal: Aging Cell

Article Title: Antiretroviral protease inhibitors induce features of cellular senescence that are reversible upon drug removal

doi: 10.1111/acel.13750

Figure Lengend Snippet: Removal of ATV/r treatment reverses senescent phenotypes in mice and improves age‐related phenotypes. Young adult mice (5‐months old) were treated with 62 mg/kg atazanavir and 21 mg/kg ritonavir in drinking water for 8 weeks. (a). Schematic of the experimental setup. (b,c). p21 Waf1 and p16 Ink4a mRNA levels in and subcutaneous fat were measured by qPCR. (d). SASP factor MMP3 mRNA levels in the subcutaneous fat tissue were measured by qPCR and normalized to Actin. (e,f). p21 Waf1 and p16 Ink4a mRNA levels in heart tissue were measured by qPCR. (g). SASP factor TIMP1 mRNA levels in the heart were measured by qPCR and normalized to Actin. (h). p16‐3MR male mice were injected with coelentarazine, and luminescence was measured at baseline, after 8 weeks of treatment, and 10 weeks post‐treatment. (i). Schematic of the experimental setup. (j). qPCR analysis for p21 Waf1 mRNA levels in heart tissue. (k). Cardiac activity was measured by echocardiography.

Article Snippet: Separated proteins were transferred to polyvinylidene fluoride membranes, which were blocked and incubated overnight at 4°C with anti‐rabbit primary antibodies (p21 WAF1 , Cell Signaling #2947, 1:1000; p‐p53, Cell Signaling #9289, 1:1000; HMGB1, abcam# ab18256, 1:2000; prelamin A, Millipore Sigma #mabt858, 1:500; β‐actin, Sigma‐Aldrich #A2228, 1:10000).

Techniques: Injection, Activity Assay