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R&D Systems rabbit antip p53

FIGURE 5 PAI-1 induced <t>p53</t> upregulation is critical for the subsequent maladaptive response. A, Western blot assessments for total and p-p53 protein levels between CMV-Con and CMV-PAI-1 populations. B-C, Histograms depicting the relative expression of p53 levels (mean ± SD) for three independent studies, n = 3. D-G, Lysates of CMV-PAI-1+Con-shRNA and CMV-PAI-1+p53-shRNA double transductants are immunoblotted for p53 (D, E; P < .001), p21 (D, F; P < .01), p-H3 (D, G; P < .01), fibronectin (H, I; P < .01), collagen-1 (H, J; P < .01). Histograms in (E-G) and (I-J) depict the relative expression (mean ± SD) for indicated proteins from the immunoblots in (D) and (H), shown as biological triplicates for three independent studies (n = 3). K, Confluent monolayers of CMV-PAI-1+Con-shRNA and CMV-PAI-1+p53- shRNA HK2 cultures are serum-starved for 6 days. Phase contrast and crystal violet images are taken on day 0 and day 6 to assess cell monolayer detachment. Scale bar = 400 µm. L, Western blot analysis for FAK and p-ERK1/2 protein levels between CMV-Con and CMV-PAI-1 cultures, with ERK2 serving as a loading control, (n = 3). *P < .05, **P < .01, ***P < .001

Rabbit Antip P53, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+antip+p53/Human+Phospho-p53+(S15)+Antibody/10__1096_slash_fj__202002652rr-32-183-188
Average 93 stars, based on 24 article reviews

rabbit antip p53 - by Bioz Stars, 2026-10

93/100 stars

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1) Product Images from "PAI‐1 induction during kidney injury promotes fibrotic epithelial dysfunction via deregulation of klotho, p53, and TGF‐β1‐receptor signaling"

Article Title: PAI‐1 induction during kidney injury promotes fibrotic epithelial dysfunction via deregulation of klotho, p53, and TGF‐β1‐receptor signaling

Journal: The FASEB Journal

doi: 10.1096/fj.202002652rr

FIGURE 5 PAI-1 induced p53 upregulation is critical for the subsequent maladaptive response. A, Western blot assessments for total and p-p53 protein levels between CMV-Con and CMV-PAI-1 populations. B-C, Histograms depicting the relative expression of p53 levels (mean ± SD) for three independent studies, n = 3. D-G, Lysates of CMV-PAI-1+Con-shRNA and CMV-PAI-1+p53-shRNA double transductants are immunoblotted for p53 (D, E; P < .001), p21 (D, F; P < .01), p-H3 (D, G; P < .01), fibronectin (H, I; P < .01), collagen-1 (H, J; P < .01). Histograms in (E-G) and (I-J) depict the relative expression (mean ± SD) for indicated proteins from the immunoblots in (D) and (H), shown as biological triplicates for three independent studies (n = 3). K, Confluent monolayers of CMV-PAI-1+Con-shRNA and CMV-PAI-1+p53- shRNA HK2 cultures are serum-starved for 6 days. Phase contrast and crystal violet images are taken on day 0 and day 6 to assess cell monolayer detachment. Scale bar = 400 µm. L, Western blot analysis for FAK and p-ERK1/2 protein levels between CMV-Con and CMV-PAI-1 cultures, with ERK2 serving as a loading control, (n = 3). *P < .05, **P < .01, ***P < .001

Figure Legend Snippet: FIGURE 5 PAI-1 induced p53 upregulation is critical for the subsequent maladaptive response. A, Western blot assessments for total and p-p53 protein levels between CMV-Con and CMV-PAI-1 populations. B-C, Histograms depicting the relative expression of p53 levels (mean ± SD) for three independent studies, n = 3. D-G, Lysates of CMV-PAI-1+Con-shRNA and CMV-PAI-1+p53-shRNA double transductants are immunoblotted for p53 (D, E; P < .001), p21 (D, F; P < .01), p-H3 (D, G; P < .01), fibronectin (H, I; P < .01), collagen-1 (H, J; P < .01). Histograms in (E-G) and (I-J) depict the relative expression (mean ± SD) for indicated proteins from the immunoblots in (D) and (H), shown as biological triplicates for three independent studies (n = 3). K, Confluent monolayers of CMV-PAI-1+Con-shRNA and CMV-PAI-1+p53- shRNA HK2 cultures are serum-starved for 6 days. Phase contrast and crystal violet images are taken on day 0 and day 6 to assess cell monolayer detachment. Scale bar = 400 µm. L, Western blot analysis for FAK and p-ERK1/2 protein levels between CMV-Con and CMV-PAI-1 cultures, with ERK2 serving as a loading control, (n = 3). *P < .05, **P < .01, ***P < .001

Techniques Used: Western Blot, Expressing, shRNA, Control

FIGURE 6 Klotho downregulation consequent to PAI-1 induction contributes to epithelial dysfunction. A-C, Immunoblot analysis of CMV- Con and CMV-PAI-1 cell lysates for PAI-1 (A, B; P < .001) and klotho (A, C; P < .01) expression. Histograms in (B-C) represent the relative expression of the indicated proteins as (mean ± SD) for three independent studies (n = 3). D-I, Protein extracts of CMV-PAI-1+CMV-Con and CMV-PAI-1+CMV-Klotho double transgenic cultures are immunoblotted for klotho (D, E; P < .001), CCN2 (D, F; P < .001), p21 (D, G; P < .01), p53 (D, H; P < .001), p-SMAD3 (D, I; P < .01). GAPDH serves as loading control. Histograms in (E-I) depict the relative levels (mean ± SD) of the indicated proteins for three separate experiments (n = 3). **P < .01, ***P < .001

Figure Legend Snippet: FIGURE 6 Klotho downregulation consequent to PAI-1 induction contributes to epithelial dysfunction. A-C, Immunoblot analysis of CMV- Con and CMV-PAI-1 cell lysates for PAI-1 (A, B; P < .001) and klotho (A, C; P < .01) expression. Histograms in (B-C) represent the relative expression of the indicated proteins as (mean ± SD) for three independent studies (n = 3). D-I, Protein extracts of CMV-PAI-1+CMV-Con and CMV-PAI-1+CMV-Klotho double transgenic cultures are immunoblotted for klotho (D, E; P < .001), CCN2 (D, F; P < .001), p21 (D, G; P < .01), p53 (D, H; P < .001), p-SMAD3 (D, I; P < .01). GAPDH serves as loading control. Histograms in (E-I) depict the relative levels (mean ± SD) of the indicated proteins for three separate experiments (n = 3). **P < .01, ***P < .001

Techniques Used: Western Blot, Expressing, Transgenic Assay, Control

FIGURE 8 Dysfunction driven by PAI-1 overexpression is independent of TGF-β1 ligand synthesis or release. A, ELISA analysis for active TGF-β1 ligand concentrations in the conditioned media isolated from serum-starved CMV-Con and CMV-PAI-1 cultures. n = 3. B-D, Cytokine protein array analysis of CMV-Con and CMV-PAI-1 conditioned media for active TGF-β1 (B), TGF-β2 (C), and TGF-β3 (D). Graphs depict the relative levels of secreted ligands (mean ± SD), n = 3. E, Immunoblot comparison of fibrotic responses in the cellular lysates of TGF-β1 stimulated or unstimulated CMV-Con and untreated CMV-PAI-1 culture extracted in parallel. F, CMV-Con HK2 cells pretreated with 20 μg/mL of TGF-β1 neutralizing antibody or 20 μg/mL IgY control antisera are stimulated with 2 ng/mL TGF-β1. Cells are harvested after 24 hours and expression of p-SMAD3, fibronectin and E-cadherin are analyzed by western blot. n = 3. G, Equally seeded CMV-PAI-1 HK2s are treated with various concentrations of TGF-β1 neutralizing antibody (0, 20, 40, 60 μg/mL) or 60 μg/mL IgY control antisera for 24 hours prior to western blot analysis of extracts for p-SMAD3, fibronectin, collagen-1, vimentin, p53 and p21, with GAPDH is serving as a loading marker. n = 3, *P < .05, **P < .01, ***P < .001, n.s., not significant. H, Western blot analysis of cell lysate extracts from CMV-PAI-1+Con shRNA and CMV-PAI-1+TGF-β1 shRNA HK2 cells for the indicated fibrotic markers. Cytokine protein array analysis of whole cell lysates (I) and conditioned media (J) used to validate TGF-β1 knockdown

Figure Legend Snippet: FIGURE 8 Dysfunction driven by PAI-1 overexpression is independent of TGF-β1 ligand synthesis or release. A, ELISA analysis for active TGF-β1 ligand concentrations in the conditioned media isolated from serum-starved CMV-Con and CMV-PAI-1 cultures. n = 3. B-D, Cytokine protein array analysis of CMV-Con and CMV-PAI-1 conditioned media for active TGF-β1 (B), TGF-β2 (C), and TGF-β3 (D). Graphs depict the relative levels of secreted ligands (mean ± SD), n = 3. E, Immunoblot comparison of fibrotic responses in the cellular lysates of TGF-β1 stimulated or unstimulated CMV-Con and untreated CMV-PAI-1 culture extracted in parallel. F, CMV-Con HK2 cells pretreated with 20 μg/mL of TGF-β1 neutralizing antibody or 20 μg/mL IgY control antisera are stimulated with 2 ng/mL TGF-β1. Cells are harvested after 24 hours and expression of p-SMAD3, fibronectin and E-cadherin are analyzed by western blot. n = 3. G, Equally seeded CMV-PAI-1 HK2s are treated with various concentrations of TGF-β1 neutralizing antibody (0, 20, 40, 60 μg/mL) or 60 μg/mL IgY control antisera for 24 hours prior to western blot analysis of extracts for p-SMAD3, fibronectin, collagen-1, vimentin, p53 and p21, with GAPDH is serving as a loading marker. n = 3, *P < .05, **P < .01, ***P < .001, n.s., not significant. H, Western blot analysis of cell lysate extracts from CMV-PAI-1+Con shRNA and CMV-PAI-1+TGF-β1 shRNA HK2 cells for the indicated fibrotic markers. Cytokine protein array analysis of whole cell lysates (I) and conditioned media (J) used to validate TGF-β1 knockdown

Techniques Used: Over Expression, Enzyme-linked Immunosorbent Assay, Isolation, Protein Array, Western Blot, Comparison, Control, Expressing, Marker, shRNA, Knockdown

FIGURE 9 Model. PAI-1 upregulation leads to downregulation of klotho, upregulation of p53, and induction of TGF-β1 receptor signaling independent of the TGF-β1 ligand, resulting in expression and secretion of fibrotic markers, downregulation of E-cadherin and upregulation of vimentin, leading to dedifferentiation, and upregulation of p21, p-H3, causing G2/M cell cycle arrest and a propensity to cell death, collectively establishing a role for PAI-1 in tubular epithelial dysfunction. Klotho regulates both p53 and SMAD3 signaling, promoting PAI-1-mediated tubular maladaptive responses

Figure Legend Snippet: FIGURE 9 Model. PAI-1 upregulation leads to downregulation of klotho, upregulation of p53, and induction of TGF-β1 receptor signaling independent of the TGF-β1 ligand, resulting in expression and secretion of fibrotic markers, downregulation of E-cadherin and upregulation of vimentin, leading to dedifferentiation, and upregulation of p21, p-H3, causing G2/M cell cycle arrest and a propensity to cell death, collectively establishing a role for PAI-1 in tubular epithelial dysfunction. Klotho regulates both p53 and SMAD3 signaling, promoting PAI-1-mediated tubular maladaptive responses

Techniques Used: Expressing

Incubation:

Article Title: PAI‐1 induction during kidney injury promotes fibrotic epithelial dysfunction via deregulation of klotho, p53, and TGF‐β1‐receptor signaling
Article Snippet: For electrophoresis, 30 μg of protein from each sample was loaded into Bio- Rad Mini- PROTEAN TGX 10% pre- cast gels and separated proteins were transferred to 0.2 μm nitrocellulose membranes and blocked in 5% milk in 0.05% Triton- X 100/ PBS before cutting membranes at the correct molecular weight. .. Overnight incubation at 4°C utilized the following primary antibodies; rabbit anti- PAI- 1 (1:3000) and rabbit anti- collagen- 1 (1:5000) as previously described,15 rat anti- klotho (1:5000; Transgenic Inc.- KM2119), rabbit anti- SMAD3 (1:1000; Cell Signaling- 9523), rabbit anti- p- Histone3 (1:1000; Cell Signaling- 9701), rabbit anti- p21 (1:1000; Cell Signaling- 2947), rabbit anti- caspase- 9 (1:1000; Cell Signaling- 9502), rabbit anti- c- caspase- 3 (1:1000; Cell Signaling- 9661), rabbit antiSnail1 (1:1000; Cell Signaling- 38798s), rabbit anti- α- SMA (1:1000; Abcam- ab32575), rabbit anti- phospho- SMAD3 (1:1000; Abcam- ab52903), rabbit anti- fibronectin (1:100,000; Abcam- ab2413), rabbit anti- LOXL2 (1:1000; Abcam- ab96233), rabbit anti- MMP- 9 (1:1000; Abcam- ab38898), rabbit antiMMP- 2 (1:1000; Abcam- ab97779), rabbit anti- VEGF- C (1:1000; Abcam- ab9546), rabbit anti- endothelin- 1 (1:1000; 18201- IBL America, Minneapolis, MN, USA), rabbit antivimentin (1:10,000; Santa Cruz- sc5565), rabbit anti- GAPDH (1:5000; Santa Cruz- sc25778), goat anti- CCN2 (1:500; Santa Cruz- sc14939), rabbit anti- TGF- βRI (1:1000; Santa Cruz- sc9048), rabbit anti- TGF- βRII (1:1000; Santa Cruz- sc220), mouse anti- p53 (1:1000; Santa Cruz- sc126), mouse anti- Ecadherin (1:1000; BD Biosciences- 610181), and rabbit antip- p53 (1:1000; AF1043- R&D systems, Minneapolis, MN, USA). .. Membranes were washed three times in 0.05% Triton- X 100/PBS before incubation with appropriate HRP- conjugated secondary antibodies (goat anti- rabbit, 31460; goat anti- mouse, 31430; both from Thermo Fisher Scientific, Waltham, MA, USA; mouse anti- goat, sc- 2354; Santa Cruz Biotechnology, Dallas, TX, USA) at a dilution of 1:5000 for 1 hour at room temperature.

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Journal: The FASEB Journal

Article Title: PAI‐1 induction during kidney injury promotes fibrotic epithelial dysfunction via deregulation of klotho, p53, and TGF‐β1‐receptor signaling

doi: 10.1096/fj.202002652rr

Figure Lengend Snippet: FIGURE 5 PAI-1 induced p53 upregulation is critical for the subsequent maladaptive response. A, Western blot assessments for total and p-p53 protein levels between CMV-Con and CMV-PAI-1 populations. B-C, Histograms depicting the relative expression of p53 levels (mean ± SD) for three independent studies, n = 3. D-G, Lysates of CMV-PAI-1+Con-shRNA and CMV-PAI-1+p53-shRNA double transductants are immunoblotted for p53 (D, E; P < .001), p21 (D, F; P < .01), p-H3 (D, G; P < .01), fibronectin (H, I; P < .01), collagen-1 (H, J; P < .01). Histograms in (E-G) and (I-J) depict the relative expression (mean ± SD) for indicated proteins from the immunoblots in (D) and (H), shown as biological triplicates for three independent studies (n = 3). K, Confluent monolayers of CMV-PAI-1+Con-shRNA and CMV-PAI-1+p53- shRNA HK2 cultures are serum-starved for 6 days. Phase contrast and crystal violet images are taken on day 0 and day 6 to assess cell monolayer detachment. Scale bar = 400 µm. L, Western blot analysis for FAK and p-ERK1/2 protein levels between CMV-Con and CMV-PAI-1 cultures, with ERK2 serving as a loading control, (n = 3). *P < .05, **P < .01, ***P < .001

Article Snippet: Overnight incubation at 4°C utilized the following primary antibodies; rabbit anti- PAI- 1 (1:3000) and rabbit anti- collagen- 1 (1:5000) as previously described,15 rat anti- klotho (1:5000; Transgenic Inc.- KM2119), rabbit anti- SMAD3 (1:1000; Cell Signaling- 9523), rabbit anti- p- Histone3 (1:1000; Cell Signaling- 9701), rabbit anti- p21 (1:1000; Cell Signaling- 2947), rabbit anti- caspase- 9 (1:1000; Cell Signaling- 9502), rabbit anti- c- caspase- 3 (1:1000; Cell Signaling- 9661), rabbit antiSnail1 (1:1000; Cell Signaling- 38798s), rabbit anti- α- SMA (1:1000; Abcam- ab32575), rabbit anti- phospho- SMAD3 (1:1000; Abcam- ab52903), rabbit anti- fibronectin (1:100,000; Abcam- ab2413), rabbit anti- LOXL2 (1:1000; Abcam- ab96233), rabbit anti- MMP- 9 (1:1000; Abcam- ab38898), rabbit antiMMP- 2 (1:1000; Abcam- ab97779), rabbit anti- VEGF- C (1:1000; Abcam- ab9546), rabbit anti- endothelin- 1 (1:1000; 18201- IBL America, Minneapolis, MN, USA), rabbit antivimentin (1:10,000; Santa Cruz- sc5565), rabbit anti- GAPDH (1:5000; Santa Cruz- sc25778), goat anti- CCN2 (1:500; Santa Cruz- sc14939), rabbit anti- TGF- βRI (1:1000; Santa Cruz- sc9048), rabbit anti- TGF- βRII (1:1000; Santa Cruz- sc220), mouse anti- p53 (1:1000; Santa Cruz- sc126), mouse anti- Ecadherin (1:1000; BD Biosciences- 610181), and rabbit antip- p53 (1:1000; AF1043- R&D systems, Minneapolis, MN, USA).

Techniques: Western Blot, Expressing, shRNA, Control

Journal: The FASEB Journal

Article Title: PAI‐1 induction during kidney injury promotes fibrotic epithelial dysfunction via deregulation of klotho, p53, and TGF‐β1‐receptor signaling

doi: 10.1096/fj.202002652rr

Figure Lengend Snippet: FIGURE 6 Klotho downregulation consequent to PAI-1 induction contributes to epithelial dysfunction. A-C, Immunoblot analysis of CMV- Con and CMV-PAI-1 cell lysates for PAI-1 (A, B; P < .001) and klotho (A, C; P < .01) expression. Histograms in (B-C) represent the relative expression of the indicated proteins as (mean ± SD) for three independent studies (n = 3). D-I, Protein extracts of CMV-PAI-1+CMV-Con and CMV-PAI-1+CMV-Klotho double transgenic cultures are immunoblotted for klotho (D, E; P < .001), CCN2 (D, F; P < .001), p21 (D, G; P < .01), p53 (D, H; P < .001), p-SMAD3 (D, I; P < .01). GAPDH serves as loading control. Histograms in (E-I) depict the relative levels (mean ± SD) of the indicated proteins for three separate experiments (n = 3). **P < .01, ***P < .001

Techniques: Western Blot, Expressing, Transgenic Assay, Control

Journal: The FASEB Journal

Article Title: PAI‐1 induction during kidney injury promotes fibrotic epithelial dysfunction via deregulation of klotho, p53, and TGF‐β1‐receptor signaling

doi: 10.1096/fj.202002652rr

Figure Lengend Snippet: FIGURE 8 Dysfunction driven by PAI-1 overexpression is independent of TGF-β1 ligand synthesis or release. A, ELISA analysis for active TGF-β1 ligand concentrations in the conditioned media isolated from serum-starved CMV-Con and CMV-PAI-1 cultures. n = 3. B-D, Cytokine protein array analysis of CMV-Con and CMV-PAI-1 conditioned media for active TGF-β1 (B), TGF-β2 (C), and TGF-β3 (D). Graphs depict the relative levels of secreted ligands (mean ± SD), n = 3. E, Immunoblot comparison of fibrotic responses in the cellular lysates of TGF-β1 stimulated or unstimulated CMV-Con and untreated CMV-PAI-1 culture extracted in parallel. F, CMV-Con HK2 cells pretreated with 20 μg/mL of TGF-β1 neutralizing antibody or 20 μg/mL IgY control antisera are stimulated with 2 ng/mL TGF-β1. Cells are harvested after 24 hours and expression of p-SMAD3, fibronectin and E-cadherin are analyzed by western blot. n = 3. G, Equally seeded CMV-PAI-1 HK2s are treated with various concentrations of TGF-β1 neutralizing antibody (0, 20, 40, 60 μg/mL) or 60 μg/mL IgY control antisera for 24 hours prior to western blot analysis of extracts for p-SMAD3, fibronectin, collagen-1, vimentin, p53 and p21, with GAPDH is serving as a loading marker. n = 3, *P < .05, **P < .01, ***P < .001, n.s., not significant. H, Western blot analysis of cell lysate extracts from CMV-PAI-1+Con shRNA and CMV-PAI-1+TGF-β1 shRNA HK2 cells for the indicated fibrotic markers. Cytokine protein array analysis of whole cell lysates (I) and conditioned media (J) used to validate TGF-β1 knockdown

Techniques: Over Expression, Enzyme-linked Immunosorbent Assay, Isolation, Protein Array, Western Blot, Comparison, Control, Expressing, Marker, shRNA, Knockdown

Journal: The FASEB Journal

Article Title: PAI‐1 induction during kidney injury promotes fibrotic epithelial dysfunction via deregulation of klotho, p53, and TGF‐β1‐receptor signaling

doi: 10.1096/fj.202002652rr

Figure Lengend Snippet: FIGURE 9 Model. PAI-1 upregulation leads to downregulation of klotho, upregulation of p53, and induction of TGF-β1 receptor signaling independent of the TGF-β1 ligand, resulting in expression and secretion of fibrotic markers, downregulation of E-cadherin and upregulation of vimentin, leading to dedifferentiation, and upregulation of p21, p-H3, causing G2/M cell cycle arrest and a propensity to cell death, collectively establishing a role for PAI-1 in tubular epithelial dysfunction. Klotho regulates both p53 and SMAD3 signaling, promoting PAI-1-mediated tubular maladaptive responses

Techniques: Expressing

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rabbit antip p53 (R&D Systems)

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1) Product Images from "PAI‐1 induction during kidney injury promotes fibrotic epithelial dysfunction via deregulation of klotho, p53, and TGF‐β1‐receptor signaling"

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