Structured Review

Santa Cruz Biotechnology p stat1
Co-Immunoprecipitation and ubiquitination of <t>P-STAT1</t> in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.
P Stat1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 99/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/p stat1/product/Santa Cruz Biotechnology
Average 99 stars, based on 44 article reviews
Price from $9.99 to $1999.99
p stat1 - by Bioz Stars, 2020-11
99/100 stars

Images

1) Product Images from "Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis"

Article Title: Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis

Journal: Surgery

doi: 10.1016/j.surg.2008.03.007

Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.
Figure Legend Snippet: Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.

Techniques Used: Immunoprecipitation, Centrifugation, Incubation

IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.
Figure Legend Snippet: IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.

Techniques Used: Ex Vivo, Mouse Assay, Expressing, Incubation, Centrifugation

Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.
Figure Legend Snippet: Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.

Techniques Used: Western Blot, Expressing

2) Product Images from "Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis"

Article Title: Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis

Journal: Surgery

doi: 10.1016/j.surg.2008.03.007

Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.
Figure Legend Snippet: Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.

Techniques Used: Immunoprecipitation, Centrifugation, Incubation

IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.
Figure Legend Snippet: IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.

Techniques Used: Ex Vivo, Mouse Assay, Expressing, Incubation, Centrifugation

Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.
Figure Legend Snippet: Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.

Techniques Used: Western Blot, Expressing

3) Product Images from "Injectable in situ cross-linking chitosan-hyaluronic acid based hydrogels for abdominal tissue regeneration"

Article Title: Injectable in situ cross-linking chitosan-hyaluronic acid based hydrogels for abdominal tissue regeneration

Journal: Scientific Reports

doi: 10.1038/s41598-017-02962-z

( A ) Expression of p-STAT1 and p-STAT6 in the regenerative tissue was determined by immunohistochemistry. The amplification was 200× . ( B ) Representative western blot images (cropped) of expression of p-STAT1, STAT1, p-STAT6 and STAT6 in the regenerative tissue. ( C ) Gray intensity of p-STAT1 and p-STAT6 was analyzed. *P
Figure Legend Snippet: ( A ) Expression of p-STAT1 and p-STAT6 in the regenerative tissue was determined by immunohistochemistry. The amplification was 200× . ( B ) Representative western blot images (cropped) of expression of p-STAT1, STAT1, p-STAT6 and STAT6 in the regenerative tissue. ( C ) Gray intensity of p-STAT1 and p-STAT6 was analyzed. *P

Techniques Used: Expressing, Immunohistochemistry, Amplification, Western Blot

4) Product Images from "LINC00963 targeting miR‐128‐3p promotes acute kidney injury process by activating JAK2/STAT1 pathway, et al. LINC00963 targeting miR‐128‐3p promotes acute kidney injury process by activating JAK2/STAT1 pathway"

Article Title: LINC00963 targeting miR‐128‐3p promotes acute kidney injury process by activating JAK2/STAT1 pathway, et al. LINC00963 targeting miR‐128‐3p promotes acute kidney injury process by activating JAK2/STAT1 pathway

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.15211

The relationship between LINC00963 and JAK2/STAT1 pathways in hypoxia‐induced HK‐2 cell model. A, The protein levels of p‐JAK2 and p‐STAT1 in different groups. B, Apoptosis analysed by flow cytometry. C, Expression of apoptosis‐related proteins by Western blot. Data are expressed as mean ± SEM, and different letters represent significant differences ( P
Figure Legend Snippet: The relationship between LINC00963 and JAK2/STAT1 pathways in hypoxia‐induced HK‐2 cell model. A, The protein levels of p‐JAK2 and p‐STAT1 in different groups. B, Apoptosis analysed by flow cytometry. C, Expression of apoptosis‐related proteins by Western blot. Data are expressed as mean ± SEM, and different letters represent significant differences ( P

Techniques Used: Flow Cytometry, Expressing, Western Blot

The relationship between LINC00963 and JAK2/STAT1 pathways on I/R‐induced AKI rat model. A, Expression of p‐JAK2 and p‐STAT1 under different conditions. B, TUNEL staining and cell apoptosis proportion of different groups. C, The protein levels of Bax and Bcl‐2. Data are expressed as mean ± SEM, and different letters represent significant differences ( P
Figure Legend Snippet: The relationship between LINC00963 and JAK2/STAT1 pathways on I/R‐induced AKI rat model. A, Expression of p‐JAK2 and p‐STAT1 under different conditions. B, TUNEL staining and cell apoptosis proportion of different groups. C, The protein levels of Bax and Bcl‐2. Data are expressed as mean ± SEM, and different letters represent significant differences ( P

Techniques Used: Expressing, TUNEL Assay, Staining

5) Product Images from "A natural mutation in the Tyk2 pseudokinase domain underlies altered susceptibility of B10.Q/J mice to infection and autoimmunity"

Article Title: A natural mutation in the Tyk2 pseudokinase domain underlies altered susceptibility of B10.Q/J mice to infection and autoimmunity

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.1930781100

B10.Q/J cells exhibit impaired cellular responses to IL-12, IFN-α, and IL-23. Impaired Stat1 phosphorylation in B10.Q/J cells after IL-12 ( a ) or IFN-α ( b ) exposure is shown. ( a ) T cell blasts derived from B10.Q/Ai, B10.Q/J, and Tyk2 – / – mice were cultured with (+) or without (–) IL-12 (50 ng/ml) for 15 min. ( b ) Lung-derived fibroblasts from B10.Q/Ai, B10.Q/J, and Tyk2 – / – mice were cultured with (+) or without (–) IFN-α (1,000 units/ml) for 15 min. Cells were analyzed for activation of Stat1 by direct Western blot with antiphosphotyrosine-Stat1 ( p -Stat1) Ab. Protein loading was controlled by reprobing with normal Stat1 Ab. ( c ) Antiviral response of B10.Q/J fibroblasts. Fibroblasts from B10.Q/Ai, B10.Q/J, and Tyk2 – / – mice were stimulated with IFN-α and cultured with vesicular stomatitis virus. After 48 h, surviving cells were quantified by crystal violet staining. ( d ) Lymphocytes from wild-type, B10.Q/J, and Tyk2 – / – mice were analyzed for Stat3 activation after 30-min stimulation with IL-12 (50 ng/ml) or IL-23 (1 μg/ml) with p -Stat3 Ab. Protein loading was controlled by reprobing with normal Stat3 Ab. a and b are representative of one of three experiments, and c and d are representative of two experiments.
Figure Legend Snippet: B10.Q/J cells exhibit impaired cellular responses to IL-12, IFN-α, and IL-23. Impaired Stat1 phosphorylation in B10.Q/J cells after IL-12 ( a ) or IFN-α ( b ) exposure is shown. ( a ) T cell blasts derived from B10.Q/Ai, B10.Q/J, and Tyk2 – / – mice were cultured with (+) or without (–) IL-12 (50 ng/ml) for 15 min. ( b ) Lung-derived fibroblasts from B10.Q/Ai, B10.Q/J, and Tyk2 – / – mice were cultured with (+) or without (–) IFN-α (1,000 units/ml) for 15 min. Cells were analyzed for activation of Stat1 by direct Western blot with antiphosphotyrosine-Stat1 ( p -Stat1) Ab. Protein loading was controlled by reprobing with normal Stat1 Ab. ( c ) Antiviral response of B10.Q/J fibroblasts. Fibroblasts from B10.Q/Ai, B10.Q/J, and Tyk2 – / – mice were stimulated with IFN-α and cultured with vesicular stomatitis virus. After 48 h, surviving cells were quantified by crystal violet staining. ( d ) Lymphocytes from wild-type, B10.Q/J, and Tyk2 – / – mice were analyzed for Stat3 activation after 30-min stimulation with IL-12 (50 ng/ml) or IL-23 (1 μg/ml) with p -Stat3 Ab. Protein loading was controlled by reprobing with normal Stat3 Ab. a and b are representative of one of three experiments, and c and d are representative of two experiments.

Techniques Used: Derivative Assay, Mouse Assay, Cell Culture, Activation Assay, Western Blot, Staining

6) Product Images from "Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation"

Article Title: Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation

Journal: Journal of Translational Medicine

doi: 10.1186/1479-5876-11-141

Dexmedetomidine inhibited the phosphorylations of JAK2, STAT1 and STAT3. Representative Western blots for the phosphorylations of JAK2, STAT1, STAT3 and total JAK2, STAT1, STAT3 expression (A) of the kidneys were detected after 48 h of renal I/R in the sham, IRI, DMSO, Atip, DEX and AG490 groups. Densitometry analysis of Western blots for the ratio of p-JAK2/JAK2 (B) , p-STAT1/STAT1 (C) and p-STAT3/STAT3 (D) . Data were represented as mean ± SEM ( n = 8). * P
Figure Legend Snippet: Dexmedetomidine inhibited the phosphorylations of JAK2, STAT1 and STAT3. Representative Western blots for the phosphorylations of JAK2, STAT1, STAT3 and total JAK2, STAT1, STAT3 expression (A) of the kidneys were detected after 48 h of renal I/R in the sham, IRI, DMSO, Atip, DEX and AG490 groups. Densitometry analysis of Western blots for the ratio of p-JAK2/JAK2 (B) , p-STAT1/STAT1 (C) and p-STAT3/STAT3 (D) . Data were represented as mean ± SEM ( n = 8). * P

Techniques Used: Western Blot, Expressing

The effect of dexmedetomidine on the expression of p-STAT1 in the kidneys following renal I/R-induced injury. Immunohistochemical staining of p-STAT1 was performed on formalin-fixed paraffin-embedded kidneys of the sham (A) , IRI (B) , DMSO (C) , Atip (D) , DEX (E) and AG490 (F) groups at 48 h following renal I/R. Bar = 50 μm. n = 8 per group.
Figure Legend Snippet: The effect of dexmedetomidine on the expression of p-STAT1 in the kidneys following renal I/R-induced injury. Immunohistochemical staining of p-STAT1 was performed on formalin-fixed paraffin-embedded kidneys of the sham (A) , IRI (B) , DMSO (C) , Atip (D) , DEX (E) and AG490 (F) groups at 48 h following renal I/R. Bar = 50 μm. n = 8 per group.

Techniques Used: Expressing, Immunohistochemistry, Staining, Formalin-fixed Paraffin-Embedded

7) Product Images from "Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation"

Article Title: Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation

Journal: Journal of Translational Medicine

doi: 10.1186/1479-5876-11-141

Dexmedetomidine inhibited the phosphorylations of JAK2, STAT1 and STAT3. Representative Western blots for the phosphorylations of JAK2, STAT1, STAT3 and total JAK2, STAT1, STAT3 expression (A) of the kidneys were detected after 48 h of renal I/R in the sham, IRI, DMSO, Atip, DEX and AG490 groups. Densitometry analysis of Western blots for the ratio of p-JAK2/JAK2 (B) , p-STAT1/STAT1 (C) and p-STAT3/STAT3 (D) . Data were represented as mean ± SEM ( n = 8). * P
Figure Legend Snippet: Dexmedetomidine inhibited the phosphorylations of JAK2, STAT1 and STAT3. Representative Western blots for the phosphorylations of JAK2, STAT1, STAT3 and total JAK2, STAT1, STAT3 expression (A) of the kidneys were detected after 48 h of renal I/R in the sham, IRI, DMSO, Atip, DEX and AG490 groups. Densitometry analysis of Western blots for the ratio of p-JAK2/JAK2 (B) , p-STAT1/STAT1 (C) and p-STAT3/STAT3 (D) . Data were represented as mean ± SEM ( n = 8). * P

Techniques Used: Western Blot, Expressing

The effect of dexmedetomidine on the expression of p-STAT1 in the kidneys following renal I/R-induced injury. Immunohistochemical staining of p-STAT1 was performed on formalin-fixed paraffin-embedded kidneys of the sham (A) , IRI (B) , DMSO (C) , Atip (D) , DEX (E) and AG490 (F) groups at 48 h following renal I/R. Bar = 50 μm. n = 8 per group.
Figure Legend Snippet: The effect of dexmedetomidine on the expression of p-STAT1 in the kidneys following renal I/R-induced injury. Immunohistochemical staining of p-STAT1 was performed on formalin-fixed paraffin-embedded kidneys of the sham (A) , IRI (B) , DMSO (C) , Atip (D) , DEX (E) and AG490 (F) groups at 48 h following renal I/R. Bar = 50 μm. n = 8 per group.

Techniques Used: Expressing, Immunohistochemistry, Staining, Formalin-fixed Paraffin-Embedded

8) Product Images from "Potential role of LMP2 as tumor-suppressor defines new targets for uterine leiomyosarcoma therapy"

Article Title: Potential role of LMP2 as tumor-suppressor defines new targets for uterine leiomyosarcoma therapy

Journal: Scientific Reports

doi: 10.1038/srep00180

Somatic JAK1 mutations prevent LMP2 expression. (a) JAK1 mutants resulted in defective activation of downstream IFN-γ pathways, as described in Table S3 . Activation of JAK1, STAT1, and IRF-1 for LMP2 expression by IFN-γ in cells expressing JAK1WT and JAK1 mutants. Protein lysates of JAK1-deficient cells expressing wild-type and JAK1 mutants were analyzed by Western blotting (W.B.) using appropriate antibodies. JAK1 and STAT1 phosphorylation after 15 minutes of stimulation was consistently increased in cells expressing JAK1WT and JAK1G986P, but phosphorylated proteins were not detected in cells expressing other JAK1 mutants. IRF-1 and LMP2 expression was markedly activated in cells expressing JAK1WT and JAK1G986P. There was no difference in the expression of JAK1, STAT1, SP1, or β-Actin among cells expressing wild-type and JAK1 mutants. (b) Defective DNA-binding activities of IRF-1 and STAT1 suggest that LMP2 expression is attributable to mutations in the catalytic domains of JAK1. DNA-binding activities of SP1 were detected in all tested samples. RT-PCR supported W.B. results. The experiments were performed three times with similar results. Direct sequencing demonstrated that the mutations detected in the catalytic sites were indeed somatic mutations (details in Supplementary Fig. S7 online).
Figure Legend Snippet: Somatic JAK1 mutations prevent LMP2 expression. (a) JAK1 mutants resulted in defective activation of downstream IFN-γ pathways, as described in Table S3 . Activation of JAK1, STAT1, and IRF-1 for LMP2 expression by IFN-γ in cells expressing JAK1WT and JAK1 mutants. Protein lysates of JAK1-deficient cells expressing wild-type and JAK1 mutants were analyzed by Western blotting (W.B.) using appropriate antibodies. JAK1 and STAT1 phosphorylation after 15 minutes of stimulation was consistently increased in cells expressing JAK1WT and JAK1G986P, but phosphorylated proteins were not detected in cells expressing other JAK1 mutants. IRF-1 and LMP2 expression was markedly activated in cells expressing JAK1WT and JAK1G986P. There was no difference in the expression of JAK1, STAT1, SP1, or β-Actin among cells expressing wild-type and JAK1 mutants. (b) Defective DNA-binding activities of IRF-1 and STAT1 suggest that LMP2 expression is attributable to mutations in the catalytic domains of JAK1. DNA-binding activities of SP1 were detected in all tested samples. RT-PCR supported W.B. results. The experiments were performed three times with similar results. Direct sequencing demonstrated that the mutations detected in the catalytic sites were indeed somatic mutations (details in Supplementary Fig. S7 online).

Techniques Used: Expressing, Activation Assay, Western Blot, Binding Assay, Reverse Transcription Polymerase Chain Reaction, Sequencing

9) Product Images from "The HER2 inhibitor TAK165 Sensitizes Human Acute Myeloid Leukemia Cells to Retinoic Acid-Induced Myeloid Differentiation by activating MEK/ERK mediated RARα/STAT1 axis"

Article Title: The HER2 inhibitor TAK165 Sensitizes Human Acute Myeloid Leukemia Cells to Retinoic Acid-Induced Myeloid Differentiation by activating MEK/ERK mediated RARα/STAT1 axis

Journal: Scientific Reports

doi: 10.1038/srep24589

Effect of TAK165 and ATRA on STAT1 activation. ( a ) Western blot analysis of p-STAT1 (Tyr701 and Ser727) and STAT1 in the HL60 and NB4 cells. The cells were treated with TAK165 (100 nM for HL60, 50 nM for NB4) and ATRA (1 μM for HL60, 2 nM for NB4) for 3 days. ( b ) The relative phosphorylation level of STAT1 at Tyr701 and Ser727 in the HL60 cells, were evaluated by p-STAT1 (Tyr701 or Ser727)/STAT1. ( c ) STAT1 expression in the NB4-scrambleand NB4-shRNA-STAT1 cells was measured by western blot. ( d ) CD11b expression in the NB4-scrambleand NB4-shRNA-STAT1 cells. The cells were treated with 50 nM TAK165 and 2 nM ATRA for 3 days. The data presented are the mean ± SD. *** p
Figure Legend Snippet: Effect of TAK165 and ATRA on STAT1 activation. ( a ) Western blot analysis of p-STAT1 (Tyr701 and Ser727) and STAT1 in the HL60 and NB4 cells. The cells were treated with TAK165 (100 nM for HL60, 50 nM for NB4) and ATRA (1 μM for HL60, 2 nM for NB4) for 3 days. ( b ) The relative phosphorylation level of STAT1 at Tyr701 and Ser727 in the HL60 cells, were evaluated by p-STAT1 (Tyr701 or Ser727)/STAT1. ( c ) STAT1 expression in the NB4-scrambleand NB4-shRNA-STAT1 cells was measured by western blot. ( d ) CD11b expression in the NB4-scrambleand NB4-shRNA-STAT1 cells. The cells were treated with 50 nM TAK165 and 2 nM ATRA for 3 days. The data presented are the mean ± SD. *** p

Techniques Used: Activation Assay, Western Blot, Expressing, shRNA

10) Product Images from "The Mechanism of Interferon Refractoriness During Hepatitis C Virus Infection and Its Reversal with a Peroxisome Proliferator-Activated Receptor α Agonist"

Article Title: The Mechanism of Interferon Refractoriness During Hepatitis C Virus Infection and Its Reversal with a Peroxisome Proliferator-Activated Receptor α Agonist

Journal: Journal of Interferon & Cytokine Research

doi: 10.1089/jir.2014.0209

STAT1 phosphorylation and ISG promoter activity is desensitized as a result of IFN-α pretreatment, and resensitized with the addition of PPARα agonist WY-14643. JFH1-infected cells were treated with 2 U/mL IFN-α for 3 weeks,
Figure Legend Snippet: STAT1 phosphorylation and ISG promoter activity is desensitized as a result of IFN-α pretreatment, and resensitized with the addition of PPARα agonist WY-14643. JFH1-infected cells were treated with 2 U/mL IFN-α for 3 weeks,

Techniques Used: Activity Assay, Infection

HCV-induced AXL expression is reduced by WY-14643. (A) The UCSC genome browser was used to identify 2 potential STAT1 binding sites (p1 and p4) in the fourth intron of AXL . (B) Binding of STAT1 to AXL was analyzed by ChIP followed by qPCR. JFH1-infected
Figure Legend Snippet: HCV-induced AXL expression is reduced by WY-14643. (A) The UCSC genome browser was used to identify 2 potential STAT1 binding sites (p1 and p4) in the fourth intron of AXL . (B) Binding of STAT1 to AXL was analyzed by ChIP followed by qPCR. JFH1-infected

Techniques Used: Expressing, Binding Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Infection

11) Product Images from "Intestinal Myofibroblasts Produce Nitric Oxide in Response to Combinatorial Cytokine Stimulation"

Article Title: Intestinal Myofibroblasts Produce Nitric Oxide in Response to Combinatorial Cytokine Stimulation

Journal: Journal of cellular physiology

doi: 10.1002/jcp.24164

Combinatorial cytokine stimulation of IMF induces iNOS expression and NO production through a signaling pathway that requires activation of Akt,STAT1, and NF-κB. A: Western blots with antibodies specific to phosphorylated Ser473 on Akt, total
Figure Legend Snippet: Combinatorial cytokine stimulation of IMF induces iNOS expression and NO production through a signaling pathway that requires activation of Akt,STAT1, and NF-κB. A: Western blots with antibodies specific to phosphorylated Ser473 on Akt, total

Techniques Used: Expressing, Activation Assay, Western Blot

12) Product Images from "Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis"

Article Title: Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis

Journal: Surgery

doi: 10.1016/j.surg.2008.03.007

Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.
Figure Legend Snippet: Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.

Techniques Used: Immunoprecipitation, Centrifugation, Incubation

IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.
Figure Legend Snippet: IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.

Techniques Used: Ex Vivo, Mouse Assay, Expressing, Incubation, Centrifugation

Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.
Figure Legend Snippet: Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.

Techniques Used: Western Blot, Expressing

13) Product Images from "Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis"

Article Title: Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis

Journal: Surgery

doi: 10.1016/j.surg.2008.03.007

Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.
Figure Legend Snippet: Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.

Techniques Used: Immunoprecipitation, Centrifugation, Incubation

IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.
Figure Legend Snippet: IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.

Techniques Used: Ex Vivo, Mouse Assay, Expressing, Incubation, Centrifugation

Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.
Figure Legend Snippet: Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.

Techniques Used: Western Blot, Expressing

14) Product Images from "Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis"

Article Title: Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis

Journal: Surgery

doi: 10.1016/j.surg.2008.03.007

Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.
Figure Legend Snippet: Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.

Techniques Used: Immunoprecipitation, Centrifugation, Incubation

IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.
Figure Legend Snippet: IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.

Techniques Used: Ex Vivo, Mouse Assay, Expressing, Incubation, Centrifugation

Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.
Figure Legend Snippet: Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.

Techniques Used: Western Blot, Expressing

15) Product Images from "Exosomes Released from M.tuberculosis Infected Cells Can Suppress IFN-? Mediated Activation of Na?ve Macrophages"

Article Title: Exosomes Released from M.tuberculosis Infected Cells Can Suppress IFN-? Mediated Activation of Na?ve Macrophages

Journal: PLoS ONE

doi: 10.1371/journal.pone.0018564

Exosomes from M.tb -infected cells do not block IFN-γ induced STAT1 phosphorylation but do inhibit IFN-γ induced expression of CIITA. BMMØ were treated +/− exosomes isolated from RAW264.7 macrophagesas described for figure 1 followed by a 30 minute incubation with IFN-γ. Cells were lysed and analyzed by Western blot for p-STAT1 (Tyr701) (A). The p44/42 MAP Kinase antibody was used as a loading control as described previously (17). BMMØ were treated with exosomes and stimulated +/− IFN-γ for 18 hours. Cells were harvested for qRT-PCR using specific primers for target gene (CIITA) and reference gene (GAPDH). Shown is the relative mRNA expression compared to untreated cells for CIITA normalized to GAPDH (B). Results are representative of two separate experiments plus standard deviation and p value
Figure Legend Snippet: Exosomes from M.tb -infected cells do not block IFN-γ induced STAT1 phosphorylation but do inhibit IFN-γ induced expression of CIITA. BMMØ were treated +/− exosomes isolated from RAW264.7 macrophagesas described for figure 1 followed by a 30 minute incubation with IFN-γ. Cells were lysed and analyzed by Western blot for p-STAT1 (Tyr701) (A). The p44/42 MAP Kinase antibody was used as a loading control as described previously (17). BMMØ were treated with exosomes and stimulated +/− IFN-γ for 18 hours. Cells were harvested for qRT-PCR using specific primers for target gene (CIITA) and reference gene (GAPDH). Shown is the relative mRNA expression compared to untreated cells for CIITA normalized to GAPDH (B). Results are representative of two separate experiments plus standard deviation and p value

Techniques Used: Infection, Blocking Assay, Expressing, Isolation, Incubation, Western Blot, Quantitative RT-PCR, Standard Deviation

16) Product Images from "Porcine bocavirus NP1 negatively regulates interferon signaling pathway by targeting the DNA-binding domain of IRF9"

Article Title: Porcine bocavirus NP1 negatively regulates interferon signaling pathway by targeting the DNA-binding domain of IRF9

Journal: Virology

doi: 10.1016/j.virol.2015.08.005

PBoV NP1 does not impair ISGF3 complex formation but inhibits its DNA-binding activity. (A) NP1 did not affect formation of the ISGF3 complex. 293T cells were transfected with FLAG-IRF9, FLAG-STAT1, FLAG-STAT2 or empty vector plasmid and HA-tagged NP1 or empty vector plasmid. At 24-h post-transfection, the cells were incubated with IFN-α for 6 h. Cells were lysed and subjected to immunoprecipitation (IP) using rabbit anti-IRF9 antibody. The whole-cell lysates and immunoprecipitation complexes were analyzed by immunoblotting using mouse anti-Flag or mouse anti-HA antibodies. (B) ChIP analysis of ISGF3 binding to the IFN-β promoter. HEK-293T cells were transfected with NP1 or empty expression vector (10 μg each). At 24 h post-transfection, the cells were incubated with IFN-α for 6 h. Cells were lysed and subjected to immunoprecipitation (IP) using rabbit anti-IRF9 antibody. ChIP assays were then performed as described in the Materials and methods section. Real-time PCR analysis of the relative binding of ISGF3 to the ISG56 promoter was performed. The results were expressed as a signal ratio, which represents the signal to the background (IgG) ratio. One of three experiments is shown.
Figure Legend Snippet: PBoV NP1 does not impair ISGF3 complex formation but inhibits its DNA-binding activity. (A) NP1 did not affect formation of the ISGF3 complex. 293T cells were transfected with FLAG-IRF9, FLAG-STAT1, FLAG-STAT2 or empty vector plasmid and HA-tagged NP1 or empty vector plasmid. At 24-h post-transfection, the cells were incubated with IFN-α for 6 h. Cells were lysed and subjected to immunoprecipitation (IP) using rabbit anti-IRF9 antibody. The whole-cell lysates and immunoprecipitation complexes were analyzed by immunoblotting using mouse anti-Flag or mouse anti-HA antibodies. (B) ChIP analysis of ISGF3 binding to the IFN-β promoter. HEK-293T cells were transfected with NP1 or empty expression vector (10 μg each). At 24 h post-transfection, the cells were incubated with IFN-α for 6 h. Cells were lysed and subjected to immunoprecipitation (IP) using rabbit anti-IRF9 antibody. ChIP assays were then performed as described in the Materials and methods section. Real-time PCR analysis of the relative binding of ISGF3 to the ISG56 promoter was performed. The results were expressed as a signal ratio, which represents the signal to the background (IgG) ratio. One of three experiments is shown.

Techniques Used: Binding Assay, Activity Assay, Transfection, Plasmid Preparation, Incubation, Immunoprecipitation, Chromatin Immunoprecipitation, Expressing, Real-time Polymerase Chain Reaction

PBoV NP1 does not degrade or prevent phosphorylation and nuclear translocation of STAT1/STAT2. (A) Effects of NP1 on STAT1/2 phosphorylation and expression. HEK-293T cells were mock-transfected or transfected with HA-tagged NP1 expression plasmid. At 24-h post-transfection, the cells were treated with IFN-α (1000 IU/ml) for 30 min. Cell lysates were collected for immunoblot analysis with antibodies directed against phosphorylated STAT1 (Y701), phosphorylated STAT2 (Y690), STAT1, STAT2, IRF9, HA, or β-actin. (B) NP1 did not prevent STAT1 or STAT2 translocation. HeLa cells were transfected with HA-tagged NP1 expression plasmid. At 24-h post-transfection, the cells were treated with IFN-α (1000 IU/ml) for 30 min. Cells were fixed and stained with mouse anti-HA specific for NP1 (green) and rabbit antibody for STAT1 or STAT2 (red). Cells were viewed under the confocal microscope (Olympus Fluoview ver. 3.1). One of three experiments is shown.
Figure Legend Snippet: PBoV NP1 does not degrade or prevent phosphorylation and nuclear translocation of STAT1/STAT2. (A) Effects of NP1 on STAT1/2 phosphorylation and expression. HEK-293T cells were mock-transfected or transfected with HA-tagged NP1 expression plasmid. At 24-h post-transfection, the cells were treated with IFN-α (1000 IU/ml) for 30 min. Cell lysates were collected for immunoblot analysis with antibodies directed against phosphorylated STAT1 (Y701), phosphorylated STAT2 (Y690), STAT1, STAT2, IRF9, HA, or β-actin. (B) NP1 did not prevent STAT1 or STAT2 translocation. HeLa cells were transfected with HA-tagged NP1 expression plasmid. At 24-h post-transfection, the cells were treated with IFN-α (1000 IU/ml) for 30 min. Cells were fixed and stained with mouse anti-HA specific for NP1 (green) and rabbit antibody for STAT1 or STAT2 (red). Cells were viewed under the confocal microscope (Olympus Fluoview ver. 3.1). One of three experiments is shown.

Techniques Used: Translocation Assay, Expressing, Transfection, Plasmid Preparation, Staining, Microscopy

17) Product Images from "LINC00963 targeting miR‐128‐3p promotes acute kidney injury process by activating JAK2/STAT1 pathway, et al. LINC00963 targeting miR‐128‐3p promotes acute kidney injury process by activating JAK2/STAT1 pathway"

Article Title: LINC00963 targeting miR‐128‐3p promotes acute kidney injury process by activating JAK2/STAT1 pathway, et al. LINC00963 targeting miR‐128‐3p promotes acute kidney injury process by activating JAK2/STAT1 pathway

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.15211

The relationship between LINC00963 and JAK2/STAT1 pathways in hypoxia‐induced HK‐2 cell model. A, The protein levels of p‐JAK2 and p‐STAT1 in different groups. B, Apoptosis analysed by flow cytometry. C, Expression of apoptosis‐related proteins by Western blot. Data are expressed as mean ± SEM, and different letters represent significant differences ( P
Figure Legend Snippet: The relationship between LINC00963 and JAK2/STAT1 pathways in hypoxia‐induced HK‐2 cell model. A, The protein levels of p‐JAK2 and p‐STAT1 in different groups. B, Apoptosis analysed by flow cytometry. C, Expression of apoptosis‐related proteins by Western blot. Data are expressed as mean ± SEM, and different letters represent significant differences ( P

Techniques Used: Flow Cytometry, Expressing, Western Blot

The relationship between LINC00963 and JAK2/STAT1 pathways on I/R‐induced AKI rat model. A, Expression of p‐JAK2 and p‐STAT1 under different conditions. B, TUNEL staining and cell apoptosis proportion of different groups. C, The protein levels of Bax and Bcl‐2. Data are expressed as mean ± SEM, and different letters represent significant differences ( P
Figure Legend Snippet: The relationship between LINC00963 and JAK2/STAT1 pathways on I/R‐induced AKI rat model. A, Expression of p‐JAK2 and p‐STAT1 under different conditions. B, TUNEL staining and cell apoptosis proportion of different groups. C, The protein levels of Bax and Bcl‐2. Data are expressed as mean ± SEM, and different letters represent significant differences ( P

Techniques Used: Expressing, TUNEL Assay, Staining

18) Product Images from "Nucleosomal dsDNA Stimulates APOL1 Expression in Human Cultured Podocytes by Activating the cGAS/IFI16-STING Signaling Pathway"

Article Title: Nucleosomal dsDNA Stimulates APOL1 Expression in Human Cultured Podocytes by Activating the cGAS/IFI16-STING Signaling Pathway

Journal: Scientific Reports

doi: 10.1038/s41598-019-51998-w

Our proposed model of nsDNA-induced APOL1 expression through engagement of the cGAS/IFI16-STING pathway in human immortalized AB8/13 podocytes. Binding of cytosolic nsDNA by cGAS and IFI16 activates STING, which subsequently activates TBK1. Activated TBK1 phosphorylates IRF3, which promotes transcription of APOL1 and IFNβ . IFNβ released from the cells (or exogenous IFNβ) binds to IFNAR. IFNAR-associated JAK1 and Tyk2 kinases then phosphorylate STAT1, which promotes transcription of APOL1 and IFI16 . A putative IFI16-mediated activation of STING is indicated by a dashed arrow. A potential cooperation between cGAS and IFI16 is indicated by a double-headed arrow. Deficient STING phosphorylation observed in cGAS- or IFI16-knockdown cells (Fig. 6 ) suggests that nsDNA-induced APOL1 expression may be mediated by a phospho-STING-independent pathway, marked by a green arrow. A dual JAK1/JAK2 inhibitor Ruxolitinib (Ruxo) suppresses STAT1 activation and thereby inhibits IFI16 expression and STAT1-mediated APOL1 expression. Since IFNAR-mediated signaling involves JAK1 but not JAK2, our model only depicts the inhibition of JAK1 by Ruxo. Ruxo does not affect IRF3-mediated APOL1 expression.
Figure Legend Snippet: Our proposed model of nsDNA-induced APOL1 expression through engagement of the cGAS/IFI16-STING pathway in human immortalized AB8/13 podocytes. Binding of cytosolic nsDNA by cGAS and IFI16 activates STING, which subsequently activates TBK1. Activated TBK1 phosphorylates IRF3, which promotes transcription of APOL1 and IFNβ . IFNβ released from the cells (or exogenous IFNβ) binds to IFNAR. IFNAR-associated JAK1 and Tyk2 kinases then phosphorylate STAT1, which promotes transcription of APOL1 and IFI16 . A putative IFI16-mediated activation of STING is indicated by a dashed arrow. A potential cooperation between cGAS and IFI16 is indicated by a double-headed arrow. Deficient STING phosphorylation observed in cGAS- or IFI16-knockdown cells (Fig. 6 ) suggests that nsDNA-induced APOL1 expression may be mediated by a phospho-STING-independent pathway, marked by a green arrow. A dual JAK1/JAK2 inhibitor Ruxolitinib (Ruxo) suppresses STAT1 activation and thereby inhibits IFI16 expression and STAT1-mediated APOL1 expression. Since IFNAR-mediated signaling involves JAK1 but not JAK2, our model only depicts the inhibition of JAK1 by Ruxo. Ruxo does not affect IRF3-mediated APOL1 expression.

Techniques Used: Expressing, Binding Assay, Activation Assay, Inhibition

nsDNA-induced APOL1 expression is partly attenuated by JAK1/JAK2 inhibitor Ruxolitinib. ( a ) Ruxolitinib (Ruxo) inhibited IFNβ-mediated phosphorylation of STAT1. Sets of AB8/13 podocytes were treated with DMSO (solvent) only (Control), treated for 2 h with 5 μM Ruxo, treated for 15 min with 10 ng ml −1 IFNβ, or pretreated with Ruxo for 2 h followed by IFNβ stimulation for 15 min (Ruxo/IFNβ). The blot images were obtained from different gels. The blot probed for P-STAT1 was re-probed for GAPDH. ( b , c ) AB8/13 podocytes were treated with Ruxo and IFNβ as indicated and subsequently transfected with 1 μg ml −1 nsDNA for 2 h ( b ) or 18 h ( c ). The IFI16/GAPDH and APOL1/GAPDH ratios in unstimulated cells (Control) were both set as 1.0. The blot images in ( b ) were obtained from different gels. The blot probed for P-IRF3 was re-probed for P-STING. Other blot images were cropped from individually probed blots. The blot images in ( c ) were obtained from different gels. The blot probed for IFI16 was re-probed for cGAS. Other blot images were cropped from individually probed blots. Full images of all blots are shown in Supplementary Fig. S7 . ( d , e ) Ruxo abolished expression of APOL1 ( d ) and IFI16 mRNA ( e ) induced by exogenous IFNβ. ( f , g ) Ruxo partially inhibited nsDNA-induced APOL1 mRNA expression ( f ) but abolished nsDNA-induced IFI16 mRNA expression ( g ). Expression of APOL1 and IFNβ mRNA was analyzed by qRT-PCR 18 h after transfection with 1 μg ml −1 nsDNA. mRNA expression was normalized to GAPDH mRNA levels. Data are expressed as means ± SEM from three biological replicates (one-way ANOVA with post-hoc Tukey test).
Figure Legend Snippet: nsDNA-induced APOL1 expression is partly attenuated by JAK1/JAK2 inhibitor Ruxolitinib. ( a ) Ruxolitinib (Ruxo) inhibited IFNβ-mediated phosphorylation of STAT1. Sets of AB8/13 podocytes were treated with DMSO (solvent) only (Control), treated for 2 h with 5 μM Ruxo, treated for 15 min with 10 ng ml −1 IFNβ, or pretreated with Ruxo for 2 h followed by IFNβ stimulation for 15 min (Ruxo/IFNβ). The blot images were obtained from different gels. The blot probed for P-STAT1 was re-probed for GAPDH. ( b , c ) AB8/13 podocytes were treated with Ruxo and IFNβ as indicated and subsequently transfected with 1 μg ml −1 nsDNA for 2 h ( b ) or 18 h ( c ). The IFI16/GAPDH and APOL1/GAPDH ratios in unstimulated cells (Control) were both set as 1.0. The blot images in ( b ) were obtained from different gels. The blot probed for P-IRF3 was re-probed for P-STING. Other blot images were cropped from individually probed blots. The blot images in ( c ) were obtained from different gels. The blot probed for IFI16 was re-probed for cGAS. Other blot images were cropped from individually probed blots. Full images of all blots are shown in Supplementary Fig. S7 . ( d , e ) Ruxo abolished expression of APOL1 ( d ) and IFI16 mRNA ( e ) induced by exogenous IFNβ. ( f , g ) Ruxo partially inhibited nsDNA-induced APOL1 mRNA expression ( f ) but abolished nsDNA-induced IFI16 mRNA expression ( g ). Expression of APOL1 and IFNβ mRNA was analyzed by qRT-PCR 18 h after transfection with 1 μg ml −1 nsDNA. mRNA expression was normalized to GAPDH mRNA levels. Data are expressed as means ± SEM from three biological replicates (one-way ANOVA with post-hoc Tukey test).

Techniques Used: Expressing, Transfection, Quantitative RT-PCR

19) Product Images from "Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation"

Article Title: Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation

Journal: Journal of Translational Medicine

doi: 10.1186/1479-5876-11-141

Dexmedetomidine inhibited the phosphorylations of JAK2, STAT1 and STAT3. Representative Western blots for the phosphorylations of JAK2, STAT1, STAT3 and total JAK2, STAT1, STAT3 expression (A) of the kidneys were detected after 48 h of renal I/R in the sham, IRI, DMSO, Atip, DEX and AG490 groups. Densitometry analysis of Western blots for the ratio of p-JAK2/JAK2 (B) , p-STAT1/STAT1 (C) and p-STAT3/STAT3 (D) . Data were represented as mean ± SEM ( n = 8). * P
Figure Legend Snippet: Dexmedetomidine inhibited the phosphorylations of JAK2, STAT1 and STAT3. Representative Western blots for the phosphorylations of JAK2, STAT1, STAT3 and total JAK2, STAT1, STAT3 expression (A) of the kidneys were detected after 48 h of renal I/R in the sham, IRI, DMSO, Atip, DEX and AG490 groups. Densitometry analysis of Western blots for the ratio of p-JAK2/JAK2 (B) , p-STAT1/STAT1 (C) and p-STAT3/STAT3 (D) . Data were represented as mean ± SEM ( n = 8). * P

Techniques Used: Western Blot, Expressing

The effect of dexmedetomidine on the expression of p-STAT1 in the kidneys following renal I/R-induced injury. Immunohistochemical staining of p-STAT1 was performed on formalin-fixed paraffin-embedded kidneys of the sham (A) , IRI (B) , DMSO (C) , Atip (D) , DEX (E) and AG490 (F) groups at 48 h following renal I/R. Bar = 50 μm. n = 8 per group.
Figure Legend Snippet: The effect of dexmedetomidine on the expression of p-STAT1 in the kidneys following renal I/R-induced injury. Immunohistochemical staining of p-STAT1 was performed on formalin-fixed paraffin-embedded kidneys of the sham (A) , IRI (B) , DMSO (C) , Atip (D) , DEX (E) and AG490 (F) groups at 48 h following renal I/R. Bar = 50 μm. n = 8 per group.

Techniques Used: Expressing, Immunohistochemistry, Staining, Formalin-fixed Paraffin-Embedded

20) Product Images from "Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis"

Article Title: Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis

Journal: Surgery

doi: 10.1016/j.surg.2008.03.007

Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.
Figure Legend Snippet: Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.

Techniques Used: Immunoprecipitation, Centrifugation, Incubation

IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.
Figure Legend Snippet: IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.

Techniques Used: Ex Vivo, Mouse Assay, Expressing, Incubation, Centrifugation

Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.
Figure Legend Snippet: Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.

Techniques Used: Western Blot, Expressing

21) Product Images from "Cell-intrinsic role for IFN-?-STAT1 signals in regulating murine Peyer patch plasmacytoid dendritic cells and conditioning an inflammatory response"

Article Title: Cell-intrinsic role for IFN-?-STAT1 signals in regulating murine Peyer patch plasmacytoid dendritic cells and conditioning an inflammatory response

Journal: Blood

doi: 10.1182/blood-2011-04-349761

Role for STAT1 in IFN-α–mediated pDC development. (A) STAT activation and expression in IFN-α–, GM-CSF-, or IFN-α + GM-CSF-treated (30 minutes) DC progenitors (lin − Flt3 + cells), determined by immunoblotting, as indicated. (B) Proportion of pDCs (top left quadrant) and cDCs (bottom right quadrant) in total BM cultures from Stat1 −/− or wild-type mice in GM-CSF or GM-CSF + IFN-α for 4 days. Results were gated on the CD11c + population (not shown); B220 and CD11b analysis is shown. n = 5. (C-D) Proportion (C) and absolute numbers (D) of pDCs and cDCs in BM and spleen of Stat1 −/− or wild-type mice treated by HGT, as indicated. Results were analyzed as indicated in panel B. n = 5. Error bars represent SEM. (E) Gene expression in D2SC/1 cells stimulated with GM-CSF, IFN-α, or both for 2 hours or left unstimulated, as indicated. Error bars represent SEM. (F) EMSAs with nuclear extracts from D2SC/1 cells stimulated with or without IFN-α for 30 minutes. Some samples contained STAT1 competitor antibody or competitor oligonucleotides, as indicated. d indicates Irf8 STAT site; m, mutated Irf8 STAT site; and s, STAT1-consensus oligonucleotide. (G) Luciferase assays in IFN-α–stimulated (IFN) or untreated (NT) D2SC/1 cells transfected with empty vector (pGL4.12) or an Irf8 reporter (pGL4.12/IRF8) with an intact (WT) or mutated (MU) STAT element, plus pMNC/mSTAT1, pMNC/mSTAT2, and phRL-TK plasmids. Error bars represent SEM. (H) ChIPs from D2SC/1 cells with or without IFN-α stimulation (1 hour) with STAT1 antibodies or IgG controls, as indicated. PCR reactions were performed with total cell lysates (input) or immunoprecipitated samples, as shown. (A-H) Results represent 3 independent experiments.
Figure Legend Snippet: Role for STAT1 in IFN-α–mediated pDC development. (A) STAT activation and expression in IFN-α–, GM-CSF-, or IFN-α + GM-CSF-treated (30 minutes) DC progenitors (lin − Flt3 + cells), determined by immunoblotting, as indicated. (B) Proportion of pDCs (top left quadrant) and cDCs (bottom right quadrant) in total BM cultures from Stat1 −/− or wild-type mice in GM-CSF or GM-CSF + IFN-α for 4 days. Results were gated on the CD11c + population (not shown); B220 and CD11b analysis is shown. n = 5. (C-D) Proportion (C) and absolute numbers (D) of pDCs and cDCs in BM and spleen of Stat1 −/− or wild-type mice treated by HGT, as indicated. Results were analyzed as indicated in panel B. n = 5. Error bars represent SEM. (E) Gene expression in D2SC/1 cells stimulated with GM-CSF, IFN-α, or both for 2 hours or left unstimulated, as indicated. Error bars represent SEM. (F) EMSAs with nuclear extracts from D2SC/1 cells stimulated with or without IFN-α for 30 minutes. Some samples contained STAT1 competitor antibody or competitor oligonucleotides, as indicated. d indicates Irf8 STAT site; m, mutated Irf8 STAT site; and s, STAT1-consensus oligonucleotide. (G) Luciferase assays in IFN-α–stimulated (IFN) or untreated (NT) D2SC/1 cells transfected with empty vector (pGL4.12) or an Irf8 reporter (pGL4.12/IRF8) with an intact (WT) or mutated (MU) STAT element, plus pMNC/mSTAT1, pMNC/mSTAT2, and phRL-TK plasmids. Error bars represent SEM. (H) ChIPs from D2SC/1 cells with or without IFN-α stimulation (1 hour) with STAT1 antibodies or IgG controls, as indicated. PCR reactions were performed with total cell lysates (input) or immunoprecipitated samples, as shown. (A-H) Results represent 3 independent experiments.

Techniques Used: Activation Assay, Expressing, Mouse Assay, Luciferase, Transfection, Plasmid Preparation, Polymerase Chain Reaction, Immunoprecipitation

Characterization of PP pDCs and their requirement for IFN-STAT1 signaling. (A) Gating strategy of PP pDCs. Surface expression of PDCA-1, B220, CCR9, CD86, and TLR4 and intracellular TLR7 and TLR9 expression in pDCs from BM, spleen, and PP. Shaded area represents isotype control staining. (B) Gene expression of Irf7 , Irf8 , and Tcf4 in pDCs from BM, spleen, and PP, determined by quantitative PCR. (C-D) Proportion (C) and absolute number (D) of PP pDCs from Stat1 −/− , Ifnar −/− , and wild-type (WT) controls. Error bars represent SEM of results from 4-8 mice. (E) PP pDC proportions in mice receiving HGT with pORF vector (NT) or pORF encoding Flt3L (10 μg), 4 days after treatment. (F) PP pDC proportions in Stat1 −/− mice at 8 days after transfer of 10 5 congenic CD45.1 + DC progenitors (BM lin − Flt3 + CD115 + c-kit int cells) intravenously. Similar results were obtained after transfer into Ifnar1 −/− mice (not shown). (G) PP pDC proportions in Stat1 −/− or Ifnar1 −/− mice 8 days after transfer of 10 5 wild-type DC progenitors (BM lin − Flt3 + CD115 + c-kit int cells) intravenously (CDP) or in animals left untreated (NT), as indicated. Some mice received Flt3L HGT 2 days before DC progenitor transfer (CDP + Flt3L) or Flt3L HGT alone (Flt3L), as shown. (B-G) Data represent 2 or 3 independent experiments. N = 4-10.
Figure Legend Snippet: Characterization of PP pDCs and their requirement for IFN-STAT1 signaling. (A) Gating strategy of PP pDCs. Surface expression of PDCA-1, B220, CCR9, CD86, and TLR4 and intracellular TLR7 and TLR9 expression in pDCs from BM, spleen, and PP. Shaded area represents isotype control staining. (B) Gene expression of Irf7 , Irf8 , and Tcf4 in pDCs from BM, spleen, and PP, determined by quantitative PCR. (C-D) Proportion (C) and absolute number (D) of PP pDCs from Stat1 −/− , Ifnar −/− , and wild-type (WT) controls. Error bars represent SEM of results from 4-8 mice. (E) PP pDC proportions in mice receiving HGT with pORF vector (NT) or pORF encoding Flt3L (10 μg), 4 days after treatment. (F) PP pDC proportions in Stat1 −/− mice at 8 days after transfer of 10 5 congenic CD45.1 + DC progenitors (BM lin − Flt3 + CD115 + c-kit int cells) intravenously. Similar results were obtained after transfer into Ifnar1 −/− mice (not shown). (G) PP pDC proportions in Stat1 −/− or Ifnar1 −/− mice 8 days after transfer of 10 5 wild-type DC progenitors (BM lin − Flt3 + CD115 + c-kit int cells) intravenously (CDP) or in animals left untreated (NT), as indicated. Some mice received Flt3L HGT 2 days before DC progenitor transfer (CDP + Flt3L) or Flt3L HGT alone (Flt3L), as shown. (B-G) Data represent 2 or 3 independent experiments. N = 4-10.

Techniques Used: Expressing, Staining, Real-time Polymerase Chain Reaction, Mouse Assay, Plasmid Preparation

22) Product Images from "IL-27 alleviates the bleomycin-induced pulmonary fibrosis by regulating the Th17 cell differentiation"

Article Title: IL-27 alleviates the bleomycin-induced pulmonary fibrosis by regulating the Th17 cell differentiation

Journal: BMC Pulmonary Medicine

doi: 10.1186/s12890-015-0012-4

The expression of JAK/STAT and TGF-β/Smad signaling pathway-related proteins in the lung tissues of the different groups on days 7 and 28. A . Western blots of JAK/STAT signaling pathway-related proteins: JAK, STAT1, STAT3, STAT5, SOCS3, and phosphorylated STAT1, STAT3, and STAT5. B . A histogram of the relative gray values shown in A . C . Western blots for the TGF-β/Smad signaling pathway-related proteins: Smad1, Smad3, TGF-βR1 and phosphorylated Smad1 and Smad3. D. A histogram of the relative gray values shown in C . Data are expressed as means ± SEM (n = 3). *p
Figure Legend Snippet: The expression of JAK/STAT and TGF-β/Smad signaling pathway-related proteins in the lung tissues of the different groups on days 7 and 28. A . Western blots of JAK/STAT signaling pathway-related proteins: JAK, STAT1, STAT3, STAT5, SOCS3, and phosphorylated STAT1, STAT3, and STAT5. B . A histogram of the relative gray values shown in A . C . Western blots for the TGF-β/Smad signaling pathway-related proteins: Smad1, Smad3, TGF-βR1 and phosphorylated Smad1 and Smad3. D. A histogram of the relative gray values shown in C . Data are expressed as means ± SEM (n = 3). *p

Techniques Used: Expressing, Western Blot

23) Product Images from "Activation of the Ras/Raf/MEK Pathway Facilitates Hepatitis C Virus Replication via Attenuation of the Interferon-JAK-STAT Pathway"

Article Title: Activation of the Ras/Raf/MEK Pathway Facilitates Hepatitis C Virus Replication via Attenuation of the Interferon-JAK-STAT Pathway

Journal: Journal of Virology

doi: 10.1128/JVI.00688-11

Proposed model for regulation of HCV replication mediated by the Ras/Raf/MEK pathway. HCV infection can activate the Ras/Raf/MEK pathway. On the other hand, activation of the Ras/Raf/MEK pathway can inhibit the JAK-STAT pathway and attenuate the expression of ISGs, such as the OAS and PKR genes, resulting in the derepression of HCV replication. This demonstrates a possible mechanism of evasion of the host antiviral system by HCV and provides a reasonable explanation for the low response to IFN therapy in HCV-infected patients. Upon HCV infection, the Ras/Raf/MEK pathway is activated and causes downregulation of IFNAR expression, reduces the phosphorylation status of STAT1 and STAT2, and lowers the expression of ISGs such as the OAS and PKR genes. All of these events result in dysfunction of the JAK-STAT pathway. By these mechanisms, HCV creates a suitable in vivo environment for its own replication.
Figure Legend Snippet: Proposed model for regulation of HCV replication mediated by the Ras/Raf/MEK pathway. HCV infection can activate the Ras/Raf/MEK pathway. On the other hand, activation of the Ras/Raf/MEK pathway can inhibit the JAK-STAT pathway and attenuate the expression of ISGs, such as the OAS and PKR genes, resulting in the derepression of HCV replication. This demonstrates a possible mechanism of evasion of the host antiviral system by HCV and provides a reasonable explanation for the low response to IFN therapy in HCV-infected patients. Upon HCV infection, the Ras/Raf/MEK pathway is activated and causes downregulation of IFNAR expression, reduces the phosphorylation status of STAT1 and STAT2, and lowers the expression of ISGs such as the OAS and PKR genes. All of these events result in dysfunction of the JAK-STAT pathway. By these mechanisms, HCV creates a suitable in vivo environment for its own replication.

Techniques Used: Infection, Activation Assay, Expressing, In Vivo

Inhibition of JAK-STAT pathway abolishes facilitation of HCV replication by the Ras/Raf/MEK pathway. (A) Cells were treated with Ruxo or DMSO for 24 h, and the phosphorylation of STAT1 was detected to confirm the inhibition of the JAK-STAT pathway by Ruxo. The expression of STAT1 and β-actin was also detected. (B) Cells infected with FL-J6/JFH5′C19Rluc2AUbi were transfected with V12 or vector, and Ruxo or DMSO was added to the culture medium 24 h before luciferase assay, as indicated. (C and D) The same experiment was performed with cells infected with JFH-1 virus. The virus titer (HCV RNA copies/ml) in the culture medium was measured (C), and core protein in the cell lysate was detected (D).
Figure Legend Snippet: Inhibition of JAK-STAT pathway abolishes facilitation of HCV replication by the Ras/Raf/MEK pathway. (A) Cells were treated with Ruxo or DMSO for 24 h, and the phosphorylation of STAT1 was detected to confirm the inhibition of the JAK-STAT pathway by Ruxo. The expression of STAT1 and β-actin was also detected. (B) Cells infected with FL-J6/JFH5′C19Rluc2AUbi were transfected with V12 or vector, and Ruxo or DMSO was added to the culture medium 24 h before luciferase assay, as indicated. (C and D) The same experiment was performed with cells infected with JFH-1 virus. The virus titer (HCV RNA copies/ml) in the culture medium was measured (C), and core protein in the cell lysate was detected (D).

Techniques Used: Inhibition, Expressing, Infection, Transfection, Plasmid Preparation, Luciferase

Analysis of the role of the Ras/Raf/MEK pathway in phosphorylation of STAT1 and STAT2 proteins. Huh7.5.1 cells were transfected with or without V12 and treated with or without U0126. Phosphorylated proteins and nonphosphorylated proteins were detected at 48 h posttransfection by Western blot analyses using the indicated antibodies.
Figure Legend Snippet: Analysis of the role of the Ras/Raf/MEK pathway in phosphorylation of STAT1 and STAT2 proteins. Huh7.5.1 cells were transfected with or without V12 and treated with or without U0126. Phosphorylated proteins and nonphosphorylated proteins were detected at 48 h posttransfection by Western blot analyses using the indicated antibodies.

Techniques Used: Transfection, Western Blot

24) Product Images from "Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis"

Article Title: Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis

Journal: Surgery

doi: 10.1016/j.surg.2008.03.007

Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.
Figure Legend Snippet: Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.

Techniques Used: Immunoprecipitation, Centrifugation, Incubation

IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.
Figure Legend Snippet: IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.

Techniques Used: Ex Vivo, Mouse Assay, Expressing, Incubation, Centrifugation

Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.
Figure Legend Snippet: Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.

Techniques Used: Western Blot, Expressing

25) Product Images from "Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation"

Article Title: Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation

Journal: Journal of Translational Medicine

doi: 10.1186/1479-5876-11-141

Dexmedetomidine inhibited the phosphorylations of JAK2, STAT1 and STAT3. Representative Western blots for the phosphorylations of JAK2, STAT1, STAT3 and total JAK2, STAT1, STAT3 expression (A) of the kidneys were detected after 48 h of renal I/R in the sham, IRI, DMSO, Atip, DEX and AG490 groups. Densitometry analysis of Western blots for the ratio of p-JAK2/JAK2 (B) , p-STAT1/STAT1 (C) and p-STAT3/STAT3 (D) . Data were represented as mean ± SEM ( n = 8). * P
Figure Legend Snippet: Dexmedetomidine inhibited the phosphorylations of JAK2, STAT1 and STAT3. Representative Western blots for the phosphorylations of JAK2, STAT1, STAT3 and total JAK2, STAT1, STAT3 expression (A) of the kidneys were detected after 48 h of renal I/R in the sham, IRI, DMSO, Atip, DEX and AG490 groups. Densitometry analysis of Western blots for the ratio of p-JAK2/JAK2 (B) , p-STAT1/STAT1 (C) and p-STAT3/STAT3 (D) . Data were represented as mean ± SEM ( n = 8). * P

Techniques Used: Western Blot, Expressing

The effect of dexmedetomidine on the expression of p-STAT1 in the kidneys following renal I/R-induced injury. Immunohistochemical staining of p-STAT1 was performed on formalin-fixed paraffin-embedded kidneys of the sham (A) , IRI (B) , DMSO (C) , Atip (D) , DEX (E) and AG490 (F) groups at 48 h following renal I/R. Bar = 50 μm. n = 8 per group.
Figure Legend Snippet: The effect of dexmedetomidine on the expression of p-STAT1 in the kidneys following renal I/R-induced injury. Immunohistochemical staining of p-STAT1 was performed on formalin-fixed paraffin-embedded kidneys of the sham (A) , IRI (B) , DMSO (C) , Atip (D) , DEX (E) and AG490 (F) groups at 48 h following renal I/R. Bar = 50 μm. n = 8 per group.

Techniques Used: Expressing, Immunohistochemistry, Staining, Formalin-fixed Paraffin-Embedded

26) Product Images from "Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation"

Article Title: Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation

Journal: Journal of Translational Medicine

doi: 10.1186/1479-5876-11-141

Dexmedetomidine inhibited the phosphorylations of JAK2, STAT1 and STAT3. Representative Western blots for the phosphorylations of JAK2, STAT1, STAT3 and total JAK2, STAT1, STAT3 expression (A) of the kidneys were detected after 48 h of renal I/R in the sham, IRI, DMSO, Atip, DEX and AG490 groups. Densitometry analysis of Western blots for the ratio of p-JAK2/JAK2 (B) , p-STAT1/STAT1 (C) and p-STAT3/STAT3 (D) . Data were represented as mean ± SEM ( n = 8). * P
Figure Legend Snippet: Dexmedetomidine inhibited the phosphorylations of JAK2, STAT1 and STAT3. Representative Western blots for the phosphorylations of JAK2, STAT1, STAT3 and total JAK2, STAT1, STAT3 expression (A) of the kidneys were detected after 48 h of renal I/R in the sham, IRI, DMSO, Atip, DEX and AG490 groups. Densitometry analysis of Western blots for the ratio of p-JAK2/JAK2 (B) , p-STAT1/STAT1 (C) and p-STAT3/STAT3 (D) . Data were represented as mean ± SEM ( n = 8). * P

Techniques Used: Western Blot, Expressing

The effect of dexmedetomidine on the expression of p-STAT1 in the kidneys following renal I/R-induced injury. Immunohistochemical staining of p-STAT1 was performed on formalin-fixed paraffin-embedded kidneys of the sham (A) , IRI (B) , DMSO (C) , Atip (D) , DEX (E) and AG490 (F) groups at 48 h following renal I/R. Bar = 50 μm. n = 8 per group.
Figure Legend Snippet: The effect of dexmedetomidine on the expression of p-STAT1 in the kidneys following renal I/R-induced injury. Immunohistochemical staining of p-STAT1 was performed on formalin-fixed paraffin-embedded kidneys of the sham (A) , IRI (B) , DMSO (C) , Atip (D) , DEX (E) and AG490 (F) groups at 48 h following renal I/R. Bar = 50 μm. n = 8 per group.

Techniques Used: Expressing, Immunohistochemistry, Staining, Formalin-fixed Paraffin-Embedded

27) Product Images from "Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation"

Article Title: Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation

Journal: Journal of Translational Medicine

doi: 10.1186/1479-5876-11-141

Dexmedetomidine inhibited the phosphorylations of JAK2, STAT1 and STAT3. Representative Western blots for the phosphorylations of JAK2, STAT1, STAT3 and total JAK2, STAT1, STAT3 expression (A) of the kidneys were detected after 48 h of renal I/R in the sham, IRI, DMSO, Atip, DEX and AG490 groups. Densitometry analysis of Western blots for the ratio of p-JAK2/JAK2 (B) , p-STAT1/STAT1 (C) and p-STAT3/STAT3 (D) . Data were represented as mean ± SEM ( n = 8). * P
Figure Legend Snippet: Dexmedetomidine inhibited the phosphorylations of JAK2, STAT1 and STAT3. Representative Western blots for the phosphorylations of JAK2, STAT1, STAT3 and total JAK2, STAT1, STAT3 expression (A) of the kidneys were detected after 48 h of renal I/R in the sham, IRI, DMSO, Atip, DEX and AG490 groups. Densitometry analysis of Western blots for the ratio of p-JAK2/JAK2 (B) , p-STAT1/STAT1 (C) and p-STAT3/STAT3 (D) . Data were represented as mean ± SEM ( n = 8). * P

Techniques Used: Western Blot, Expressing

The effect of dexmedetomidine on the expression of p-STAT1 in the kidneys following renal I/R-induced injury. Immunohistochemical staining of p-STAT1 was performed on formalin-fixed paraffin-embedded kidneys of the sham (A) , IRI (B) , DMSO (C) , Atip (D) , DEX (E) and AG490 (F) groups at 48 h following renal I/R. Bar = 50 μm. n = 8 per group.
Figure Legend Snippet: The effect of dexmedetomidine on the expression of p-STAT1 in the kidneys following renal I/R-induced injury. Immunohistochemical staining of p-STAT1 was performed on formalin-fixed paraffin-embedded kidneys of the sham (A) , IRI (B) , DMSO (C) , Atip (D) , DEX (E) and AG490 (F) groups at 48 h following renal I/R. Bar = 50 μm. n = 8 per group.

Techniques Used: Expressing, Immunohistochemistry, Staining, Formalin-fixed Paraffin-Embedded

28) Product Images from "Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation"

Article Title: Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation

Journal: Journal of Translational Medicine

doi: 10.1186/1479-5876-11-141

Dexmedetomidine inhibited the phosphorylations of JAK2, STAT1 and STAT3. Representative Western blots for the phosphorylations of JAK2, STAT1, STAT3 and total JAK2, STAT1, STAT3 expression (A) of the kidneys were detected after 48 h of renal I/R in the sham, IRI, DMSO, Atip, DEX and AG490 groups. Densitometry analysis of Western blots for the ratio of p-JAK2/JAK2 (B) , p-STAT1/STAT1 (C) and p-STAT3/STAT3 (D) . Data were represented as mean ± SEM ( n = 8). * P
Figure Legend Snippet: Dexmedetomidine inhibited the phosphorylations of JAK2, STAT1 and STAT3. Representative Western blots for the phosphorylations of JAK2, STAT1, STAT3 and total JAK2, STAT1, STAT3 expression (A) of the kidneys were detected after 48 h of renal I/R in the sham, IRI, DMSO, Atip, DEX and AG490 groups. Densitometry analysis of Western blots for the ratio of p-JAK2/JAK2 (B) , p-STAT1/STAT1 (C) and p-STAT3/STAT3 (D) . Data were represented as mean ± SEM ( n = 8). * P

Techniques Used: Western Blot, Expressing

The effect of dexmedetomidine on the expression of p-STAT1 in the kidneys following renal I/R-induced injury. Immunohistochemical staining of p-STAT1 was performed on formalin-fixed paraffin-embedded kidneys of the sham (A) , IRI (B) , DMSO (C) , Atip (D) , DEX (E) and AG490 (F) groups at 48 h following renal I/R. Bar = 50 μm. n = 8 per group.
Figure Legend Snippet: The effect of dexmedetomidine on the expression of p-STAT1 in the kidneys following renal I/R-induced injury. Immunohistochemical staining of p-STAT1 was performed on formalin-fixed paraffin-embedded kidneys of the sham (A) , IRI (B) , DMSO (C) , Atip (D) , DEX (E) and AG490 (F) groups at 48 h following renal I/R. Bar = 50 μm. n = 8 per group.

Techniques Used: Expressing, Immunohistochemistry, Staining, Formalin-fixed Paraffin-Embedded

29) Product Images from "Effects and mechanisms of vitamin A and vitamin E on the levels of serum leptin and other related cytokines in rats with rheumatoid arthritis"

Article Title: Effects and mechanisms of vitamin A and vitamin E on the levels of serum leptin and other related cytokines in rats with rheumatoid arthritis

Journal: Experimental and Therapeutic Medicine

doi: 10.3892/etm.2014.1777

Effects of VitA and VitE on the p-STAT1 protein expression levels in CIA model rats. (A) Western blot analysis of p-STAT1 protein levels. (B) Statistical analysis of the p-STAT1 expression levels. The relative quantification of the p-STAT1 protein levels is expressed as a ratio of the target protein to β-actin. Values were normalized with respect to β-actin and expressed as a percentage of the control. Each value represents the mean ± SD, ## P
Figure Legend Snippet: Effects of VitA and VitE on the p-STAT1 protein expression levels in CIA model rats. (A) Western blot analysis of p-STAT1 protein levels. (B) Statistical analysis of the p-STAT1 expression levels. The relative quantification of the p-STAT1 protein levels is expressed as a ratio of the target protein to β-actin. Values were normalized with respect to β-actin and expressed as a percentage of the control. Each value represents the mean ± SD, ## P

Techniques Used: Expressing, Western Blot

30) Product Images from "Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis"

Article Title: Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis

Journal: Surgery

doi: 10.1016/j.surg.2008.03.007

Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.
Figure Legend Snippet: Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.

Techniques Used: Immunoprecipitation, Centrifugation, Incubation

IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.
Figure Legend Snippet: IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.

Techniques Used: Ex Vivo, Mouse Assay, Expressing, Incubation, Centrifugation

Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.
Figure Legend Snippet: Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.

Techniques Used: Western Blot, Expressing

31) Product Images from "Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis"

Article Title: Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis

Journal: Surgery

doi: 10.1016/j.surg.2008.03.007

Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.
Figure Legend Snippet: Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.

Techniques Used: Immunoprecipitation, Centrifugation, Incubation

IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.
Figure Legend Snippet: IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.

Techniques Used: Ex Vivo, Mouse Assay, Expressing, Incubation, Centrifugation

Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.
Figure Legend Snippet: Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.

Techniques Used: Western Blot, Expressing

32) Product Images from "Hepatitis C Virus Driven AXL Expression Suppresses the Hepatic Type I Interferon Response"

Article Title: Hepatitis C Virus Driven AXL Expression Suppresses the Hepatic Type I Interferon Response

Journal: PLoS ONE

doi: 10.1371/journal.pone.0136227

AXL overexpression dampens the response to IFNα. Stable transfection of Huh-7 cells with PCMV6-AXL was confirmed by western blot, using antibodies against AXL or the fusion Myc tag (A). Huh-7 cells overexpressing AXL induced a stronger but more transient phosphorylation of STAT1 (B), with increased phosphorylation at 15 and 30 minutes post-IFN (50 u/ml), but a strong decrease at 1 h (2 replicates). ISRE and NFκB promoter activation was decreased almost 2 fold at 8 h post-IFN treatment in cells overexpressing AXL, while GAS activation were increased 2 fold at 4 h post-IFN treatment (C). No effect on AP-1 promoter activation was observed (* p
Figure Legend Snippet: AXL overexpression dampens the response to IFNα. Stable transfection of Huh-7 cells with PCMV6-AXL was confirmed by western blot, using antibodies against AXL or the fusion Myc tag (A). Huh-7 cells overexpressing AXL induced a stronger but more transient phosphorylation of STAT1 (B), with increased phosphorylation at 15 and 30 minutes post-IFN (50 u/ml), but a strong decrease at 1 h (2 replicates). ISRE and NFκB promoter activation was decreased almost 2 fold at 8 h post-IFN treatment in cells overexpressing AXL, while GAS activation were increased 2 fold at 4 h post-IFN treatment (C). No effect on AP-1 promoter activation was observed (* p

Techniques Used: Over Expression, Stable Transfection, Western Blot, Activation Assay

AXL expression is mediated by multiple innate immune signalling pathways. To identify inflammatory transcription factor binding sites within the AXL promoter/enhancer region that may mediate AXL expression, the UCSC genome browser was interrogated for experimental ChIP binding sites (A). Furthermore, the ECR browser and rVista were used to identify evolutionarily conserved regions and to predict transcription factor binding sites, respectively. Huh-7 cells were treated with a range of antiviral cytokines for 24 h then AXL expression measured by qPCR (B). IFNs α, β, and λ up-regulated AXL more potently than pro-inflammatory cytokines IFNγ and IL6 (average of two biological replicates in duplicate for each). (C) To inhibit potential signalling components that induce AXL , Huh-7 cells were treated with 10nM siRNA against STAT1/STAT3 or chemical inhibitors of JNK (SP600125) or NFκB (BAY11-7082) for 24 h. Inhibition of STAT1, STAT3, JNK (SP600125) and NFκB (BAY 11–7082) all significantly reduced HCV induced AXL expression (* p
Figure Legend Snippet: AXL expression is mediated by multiple innate immune signalling pathways. To identify inflammatory transcription factor binding sites within the AXL promoter/enhancer region that may mediate AXL expression, the UCSC genome browser was interrogated for experimental ChIP binding sites (A). Furthermore, the ECR browser and rVista were used to identify evolutionarily conserved regions and to predict transcription factor binding sites, respectively. Huh-7 cells were treated with a range of antiviral cytokines for 24 h then AXL expression measured by qPCR (B). IFNs α, β, and λ up-regulated AXL more potently than pro-inflammatory cytokines IFNγ and IL6 (average of two biological replicates in duplicate for each). (C) To inhibit potential signalling components that induce AXL , Huh-7 cells were treated with 10nM siRNA against STAT1/STAT3 or chemical inhibitors of JNK (SP600125) or NFκB (BAY11-7082) for 24 h. Inhibition of STAT1, STAT3, JNK (SP600125) and NFκB (BAY 11–7082) all significantly reduced HCV induced AXL expression (* p

Techniques Used: Expressing, Binding Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Inhibition

33) Product Images from "LINC00963 targeting miR‐128‐3p promotes acute kidney injury process by activating JAK2/STAT1 pathway, et al. LINC00963 targeting miR‐128‐3p promotes acute kidney injury process by activating JAK2/STAT1 pathway"

Article Title: LINC00963 targeting miR‐128‐3p promotes acute kidney injury process by activating JAK2/STAT1 pathway, et al. LINC00963 targeting miR‐128‐3p promotes acute kidney injury process by activating JAK2/STAT1 pathway

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.15211

The relationship between LINC00963 and JAK2/STAT1 pathways in hypoxia‐induced HK‐2 cell model. A, The protein levels of p‐JAK2 and p‐STAT1 in different groups. B, Apoptosis analysed by flow cytometry. C, Expression of apoptosis‐related proteins by Western blot. Data are expressed as mean ± SEM, and different letters represent significant differences ( P
Figure Legend Snippet: The relationship between LINC00963 and JAK2/STAT1 pathways in hypoxia‐induced HK‐2 cell model. A, The protein levels of p‐JAK2 and p‐STAT1 in different groups. B, Apoptosis analysed by flow cytometry. C, Expression of apoptosis‐related proteins by Western blot. Data are expressed as mean ± SEM, and different letters represent significant differences ( P

Techniques Used: Flow Cytometry, Expressing, Western Blot

The relationship between LINC00963 and JAK2/STAT1 pathways on I/R‐induced AKI rat model. A, Expression of p‐JAK2 and p‐STAT1 under different conditions. B, TUNEL staining and cell apoptosis proportion of different groups. C, The protein levels of Bax and Bcl‐2. Data are expressed as mean ± SEM, and different letters represent significant differences ( P
Figure Legend Snippet: The relationship between LINC00963 and JAK2/STAT1 pathways on I/R‐induced AKI rat model. A, Expression of p‐JAK2 and p‐STAT1 under different conditions. B, TUNEL staining and cell apoptosis proportion of different groups. C, The protein levels of Bax and Bcl‐2. Data are expressed as mean ± SEM, and different letters represent significant differences ( P

Techniques Used: Expressing, TUNEL Assay, Staining

34) Product Images from "Identification of human thioredoxin as a novel IFN-gamma-induced factor: Mechanism of induction and its role in cytokine production"

Article Title: Identification of human thioredoxin as a novel IFN-gamma-induced factor: Mechanism of induction and its role in cytokine production

Journal: BMC Immunology

doi: 10.1186/1471-2172-9-64

Analysis of signaling pathways activated by IFN-γ and the effects of signaling inhibitors on thioredoxin gene expression . A. Analysis of Jak, Akt, and MAPK pathways during IFN-γ signal transduction in THP1 cells . THP1 cells (1 × 10 6 ) were treated with media alone or IFN-γ under serum-free conditions for the indicated times. The total cell lysates were then prepared and analyzed by Western blot to determine the activation status of Jak1/Stat1, Akt, and MAPKs as described in the text. B. Effects of signaling inhibitors on IFN-γ-induced thioredoxin gene expression . THP1 cells (1 × 10 6 ) were pretreated with LY294002 (20 μM), PD98059 (50 μM), SP600125 (10 μM), AG490 (10 μM), PDTC (100 μM) or SB203580 (20 μM) for 1 h and then cultured for an additional 24 h in the presence or absence of IFN-γ under serum-free conditions. The total RNA was isolated and the thioredoxin mRNA levels were then analyzed by RT-PCR. C. Analysis of activation and nuclear translocation of Stat1, NF-κB, and AP-1 induced by IFN-γ in THP1 cells . Cells (5 × 10 6 ) were treated with media alone or IFN-γ (10 ng/ml) under serum-free conditions for the indicated times. The cells were then harvested and used to prepare cytoplasmic or nuclear extracts. The extracts were then subjected to Western blot to determine the activation status of Stat1, c-Jun, c-fos or NF-κB, using antibodies to anti-phosphotyrosine Stat1, anti-Stat1, c-Jun, c-fos and NF-κB (p65). As internal controls, Cox-4 and H2A were used for cytosolic and nuclear marker proteins, respectively.
Figure Legend Snippet: Analysis of signaling pathways activated by IFN-γ and the effects of signaling inhibitors on thioredoxin gene expression . A. Analysis of Jak, Akt, and MAPK pathways during IFN-γ signal transduction in THP1 cells . THP1 cells (1 × 10 6 ) were treated with media alone or IFN-γ under serum-free conditions for the indicated times. The total cell lysates were then prepared and analyzed by Western blot to determine the activation status of Jak1/Stat1, Akt, and MAPKs as described in the text. B. Effects of signaling inhibitors on IFN-γ-induced thioredoxin gene expression . THP1 cells (1 × 10 6 ) were pretreated with LY294002 (20 μM), PD98059 (50 μM), SP600125 (10 μM), AG490 (10 μM), PDTC (100 μM) or SB203580 (20 μM) for 1 h and then cultured for an additional 24 h in the presence or absence of IFN-γ under serum-free conditions. The total RNA was isolated and the thioredoxin mRNA levels were then analyzed by RT-PCR. C. Analysis of activation and nuclear translocation of Stat1, NF-κB, and AP-1 induced by IFN-γ in THP1 cells . Cells (5 × 10 6 ) were treated with media alone or IFN-γ (10 ng/ml) under serum-free conditions for the indicated times. The cells were then harvested and used to prepare cytoplasmic or nuclear extracts. The extracts were then subjected to Western blot to determine the activation status of Stat1, c-Jun, c-fos or NF-κB, using antibodies to anti-phosphotyrosine Stat1, anti-Stat1, c-Jun, c-fos and NF-κB (p65). As internal controls, Cox-4 and H2A were used for cytosolic and nuclear marker proteins, respectively.

Techniques Used: Expressing, Transduction, Western Blot, Activation Assay, Cell Culture, Isolation, Reverse Transcription Polymerase Chain Reaction, Translocation Assay, Marker

35) Product Images from "The Anti-Inflammatory Effects of Shinbaro3 Is Mediated by Downregulation of the TLR4 Signalling Pathway in LPS-Stimulated RAW 264.7 Macrophages"

Article Title: The Anti-Inflammatory Effects of Shinbaro3 Is Mediated by Downregulation of the TLR4 Signalling Pathway in LPS-Stimulated RAW 264.7 Macrophages

Journal: Mediators of Inflammation

doi: 10.1155/2018/4514329

Effects of Shinbaro3 on the IRF3/STAT1 signalling pathway in LPS-stimulated RAW 264.7 cells. RAW 264.7 cells were treated with LPS (1 μ g/mL) and Shinbaro3 (150, 300, and 450 μ g/mL) for 4 h. The expression levels of (a) IRF3, STAT1, JAK1, and their phosphorylated forms were detected with specific antibodies. β -Actin was used as an internal control. The data are representative of three separate experiments. (b) Protein expression of INF- β was assessed in RAW 264.7 cells under the above condition. (c) RAW 264.7 cells were treated with LPS (1 μ g/mL) and Shinbaro3 (150, 300, and 450 μ g/mL) for 6 h. Expression of IFN- β was investigated with real-time RT-PCR as described in Materials and Methods. (d) RAW 264.7 cells were stimulated with Shinbaro3 (150, 300, and 450 μ g/mL) in presence of IFN- β (100 U/mL) for 4 h. Protein expression of JAK1 and STAT1 and their phosphorylated forms were detected. The data are expressed as the mean ± SD ( n = 3). ∗ p
Figure Legend Snippet: Effects of Shinbaro3 on the IRF3/STAT1 signalling pathway in LPS-stimulated RAW 264.7 cells. RAW 264.7 cells were treated with LPS (1 μ g/mL) and Shinbaro3 (150, 300, and 450 μ g/mL) for 4 h. The expression levels of (a) IRF3, STAT1, JAK1, and their phosphorylated forms were detected with specific antibodies. β -Actin was used as an internal control. The data are representative of three separate experiments. (b) Protein expression of INF- β was assessed in RAW 264.7 cells under the above condition. (c) RAW 264.7 cells were treated with LPS (1 μ g/mL) and Shinbaro3 (150, 300, and 450 μ g/mL) for 6 h. Expression of IFN- β was investigated with real-time RT-PCR as described in Materials and Methods. (d) RAW 264.7 cells were stimulated with Shinbaro3 (150, 300, and 450 μ g/mL) in presence of IFN- β (100 U/mL) for 4 h. Protein expression of JAK1 and STAT1 and their phosphorylated forms were detected. The data are expressed as the mean ± SD ( n = 3). ∗ p

Techniques Used: Expressing, Quantitative RT-PCR

36) Product Images from "Bilobalide Alleviated Dextran Sulfate Sodium-Induced Experimental Colitis by Inhibiting M1 Macrophage Polarization Through the NF-κB Signaling Pathway"

Article Title: Bilobalide Alleviated Dextran Sulfate Sodium-Induced Experimental Colitis by Inhibiting M1 Macrophage Polarization Through the NF-κB Signaling Pathway

Journal: Frontiers in Pharmacology

doi: 10.3389/fphar.2020.00718

Bilobalide inhibited NF-κB signaling which hampered differentiation of M1 macrophage. (A) Bone marrow-derived macrophages (BMDMs) were treated with 1, 3, and 10 μM bilobalide in the presence of 10 ng/ml lipopolysaccharide (LPS) and 10 ng/ml interferon-gamma (IFN-γ) (M1) for 30 min. The expression of MyD88, TRAF6, p-STAT1, STAT1, p-p65, and p65 were determined by western blot. (B) BMDMs were treated with 10 μM bilobalide in the presence of 10 ng/ml LPS and 10 ng/ml IFN-γ (M1) for 30 min. The cytosol and nuclei fraction were isolated and the level of p65 was determined by western blot. (C) The BMDMs were cultured as described above and stained with p65. The nuclei were stained with 4′, 6-diamidino-2-phenylindole (DAPI) (blue). Scale bar: 10 μm.
Figure Legend Snippet: Bilobalide inhibited NF-κB signaling which hampered differentiation of M1 macrophage. (A) Bone marrow-derived macrophages (BMDMs) were treated with 1, 3, and 10 μM bilobalide in the presence of 10 ng/ml lipopolysaccharide (LPS) and 10 ng/ml interferon-gamma (IFN-γ) (M1) for 30 min. The expression of MyD88, TRAF6, p-STAT1, STAT1, p-p65, and p65 were determined by western blot. (B) BMDMs were treated with 10 μM bilobalide in the presence of 10 ng/ml LPS and 10 ng/ml IFN-γ (M1) for 30 min. The cytosol and nuclei fraction were isolated and the level of p65 was determined by western blot. (C) The BMDMs were cultured as described above and stained with p65. The nuclei were stained with 4′, 6-diamidino-2-phenylindole (DAPI) (blue). Scale bar: 10 μm.

Techniques Used: Derivative Assay, Expressing, Western Blot, Isolation, Cell Culture, Staining

37) Product Images from "B7-H3 in Medulloblastoma-Derived Exosomes; A Novel Tumorigenic Role"

Article Title: B7-H3 in Medulloblastoma-Derived Exosomes; A Novel Tumorigenic Role

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms21197050

B7-H3 expression can be induced via MB-derived exosomes in recipient cells: ( A ) Western blot of isolated exosomes (5 µg/lane) from D283 cells showing increased B7-H3 presence in B7-H3 OE transfected cell-derived exosomes compared to control. CD63 used as exosomal loading control. ( B ) F-actin staining (red) of D283, D425, and D458 cells incubated with D283-derived B7-H3 OE exosomes (green). Briefly, 15 µg of exosomes (5 µg/well) were incubated with 0.5 µL of Calcein AM in SFM for 30 min in an Eppendorf tube and then overlaid on a 3-well chambered slide (ibidi, Fitchburg, WI) with 1 mL media containing D283, D425, and D458 cells for 6 h. Yellow arrows indicate likely areas of stained exosomes fusing with target cells. ( C ) Western blot of HMECs incubated (24 h) in SFM alone, with D283 control exosomes, or D283 B7-H3 OE exosomes showing increased B7-H3 expression intracellularly. Actin was used as a loading control. The B7-H3 and Actin blots shown here are cropped using PowerPoint for clear representation. Lane 3 from the original blots of both B7-H3 and Actin were cropped from the figure as it is not relevant to this study. Lane 4, which represents exosomes from B7-H3 overexpression (EXO_B7-H3 OE), is now represented as a single lane with actin as a loading control. The corresponding uncropped full-length blot is included in Figure S3 . ( D ) Western blot of UW228 cells incubated (6 h) with SFM alone, D283 control exosomes, or D283 B7-H3 OE exosomes showing increased B7-H3 expression along with decreased STAT1 phosphorylation. Actin was used as a loading control.
Figure Legend Snippet: B7-H3 expression can be induced via MB-derived exosomes in recipient cells: ( A ) Western blot of isolated exosomes (5 µg/lane) from D283 cells showing increased B7-H3 presence in B7-H3 OE transfected cell-derived exosomes compared to control. CD63 used as exosomal loading control. ( B ) F-actin staining (red) of D283, D425, and D458 cells incubated with D283-derived B7-H3 OE exosomes (green). Briefly, 15 µg of exosomes (5 µg/well) were incubated with 0.5 µL of Calcein AM in SFM for 30 min in an Eppendorf tube and then overlaid on a 3-well chambered slide (ibidi, Fitchburg, WI) with 1 mL media containing D283, D425, and D458 cells for 6 h. Yellow arrows indicate likely areas of stained exosomes fusing with target cells. ( C ) Western blot of HMECs incubated (24 h) in SFM alone, with D283 control exosomes, or D283 B7-H3 OE exosomes showing increased B7-H3 expression intracellularly. Actin was used as a loading control. The B7-H3 and Actin blots shown here are cropped using PowerPoint for clear representation. Lane 3 from the original blots of both B7-H3 and Actin were cropped from the figure as it is not relevant to this study. Lane 4, which represents exosomes from B7-H3 overexpression (EXO_B7-H3 OE), is now represented as a single lane with actin as a loading control. The corresponding uncropped full-length blot is included in Figure S3 . ( D ) Western blot of UW228 cells incubated (6 h) with SFM alone, D283 control exosomes, or D283 B7-H3 OE exosomes showing increased B7-H3 expression along with decreased STAT1 phosphorylation. Actin was used as a loading control.

Techniques Used: Expressing, Derivative Assay, Western Blot, Isolation, Transfection, Staining, Incubation, Over Expression

38) Product Images from "Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation"

Article Title: Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation

Journal: Journal of Translational Medicine

doi: 10.1186/1479-5876-11-141

Dexmedetomidine inhibited the phosphorylations of JAK2, STAT1 and STAT3. Representative Western blots for the phosphorylations of JAK2, STAT1, STAT3 and total JAK2, STAT1, STAT3 expression (A) of the kidneys were detected after 48 h of renal I/R in the sham, IRI, DMSO, Atip, DEX and AG490 groups. Densitometry analysis of Western blots for the ratio of p-JAK2/JAK2 (B) , p-STAT1/STAT1 (C) and p-STAT3/STAT3 (D) . Data were represented as mean ± SEM ( n = 8). * P
Figure Legend Snippet: Dexmedetomidine inhibited the phosphorylations of JAK2, STAT1 and STAT3. Representative Western blots for the phosphorylations of JAK2, STAT1, STAT3 and total JAK2, STAT1, STAT3 expression (A) of the kidneys were detected after 48 h of renal I/R in the sham, IRI, DMSO, Atip, DEX and AG490 groups. Densitometry analysis of Western blots for the ratio of p-JAK2/JAK2 (B) , p-STAT1/STAT1 (C) and p-STAT3/STAT3 (D) . Data were represented as mean ± SEM ( n = 8). * P

Techniques Used: Western Blot, Expressing

The effect of dexmedetomidine on the expression of p-STAT1 in the kidneys following renal I/R-induced injury. Immunohistochemical staining of p-STAT1 was performed on formalin-fixed paraffin-embedded kidneys of the sham (A) , IRI (B) , DMSO (C) , Atip (D) , DEX (E) and AG490 (F) groups at 48 h following renal I/R. Bar = 50 μm. n = 8 per group.
Figure Legend Snippet: The effect of dexmedetomidine on the expression of p-STAT1 in the kidneys following renal I/R-induced injury. Immunohistochemical staining of p-STAT1 was performed on formalin-fixed paraffin-embedded kidneys of the sham (A) , IRI (B) , DMSO (C) , Atip (D) , DEX (E) and AG490 (F) groups at 48 h following renal I/R. Bar = 50 μm. n = 8 per group.

Techniques Used: Expressing, Immunohistochemistry, Staining, Formalin-fixed Paraffin-Embedded

39) Product Images from "microRNA-200a silencing protects neural stem cells against cerebral ischemia/reperfusion injury"

Article Title: microRNA-200a silencing protects neural stem cells against cerebral ischemia/reperfusion injury

Journal: PLoS ONE

doi: 10.1371/journal.pone.0172178

miR-200a inhibits c-Myc involved in STAT and MAPK signaling pathways. (A) and (B) NSCs were transfected with vector or shRNA against miR-200a, and then the miR-200a-silenced cells were treated with or without Nifuroxazide. p-STAT1, STAT1, p-STAT3, STAT3 and c-Myc expression levels were determined by Western blotting. (C) and (D) NSCs were transfected with vector or shRNA against miR-200a, and then the miR-200a-silenced cells were treated with or without BIRB 796. p-MAPK, MAPK and c-Myc expression levels were also detected by Western blotting. ** P
Figure Legend Snippet: miR-200a inhibits c-Myc involved in STAT and MAPK signaling pathways. (A) and (B) NSCs were transfected with vector or shRNA against miR-200a, and then the miR-200a-silenced cells were treated with or without Nifuroxazide. p-STAT1, STAT1, p-STAT3, STAT3 and c-Myc expression levels were determined by Western blotting. (C) and (D) NSCs were transfected with vector or shRNA against miR-200a, and then the miR-200a-silenced cells were treated with or without BIRB 796. p-MAPK, MAPK and c-Myc expression levels were also detected by Western blotting. ** P

Techniques Used: Transfection, Plasmid Preparation, shRNA, Expressing, Western Blot

40) Product Images from "Identification of 5-Methoxy-2-(Diformylmethylidene)-3,3-Dimethylindole as an Anti-Influenza A Virus Agent"

Article Title: Identification of 5-Methoxy-2-(Diformylmethylidene)-3,3-Dimethylindole as an Anti-Influenza A Virus Agent

Journal: PLoS ONE

doi: 10.1371/journal.pone.0170352

526A inhibits IAV-induced IRF3 and STAT1 activation. The A549-PB1 cells were treated with various concentrations of 526A as indicated for four hours followed by PR8-PB1flank-eGFP IAV infection at an MOI of 1.0 for twelve hours. Whole cell extracts were prepared and immunoblotted with antibodies against PARP, p-IRF3 and p-STAT1.
Figure Legend Snippet: 526A inhibits IAV-induced IRF3 and STAT1 activation. The A549-PB1 cells were treated with various concentrations of 526A as indicated for four hours followed by PR8-PB1flank-eGFP IAV infection at an MOI of 1.0 for twelve hours. Whole cell extracts were prepared and immunoblotted with antibodies against PARP, p-IRF3 and p-STAT1.

Techniques Used: Activation Assay, Infection

Related Articles

Immunoprecipitation:

Article Title: Stat1 Acetylation Inhibits Inducible Nitric Oxide Synthase Expression in Interferon-γ Treated RAW264.7 Murine Macrophages
Article Snippet: .. Purified chromatin was immunoprecipitated using 10 µg of anti-NF-κB p65 (Santa Cruz Biotechnology, Santa Cruz, CA), anti-Stat1 (Santa Cruz Biotechnology, Santa Cruz, CA) or 5 µl of rabbit nonimmune serum; eluted DNA fragments were purified to serve as templates. .. The input fraction corresponded to 0.1 and 0.05% of the chromatin solution before immunoprecipitation.

Incubation:

Article Title: Altered Levels of STAT1 and STAT3 Influence the Neuronal Response to Interferon Gamma
Article Snippet: .. After three washes in PBS-T (5 min each), the blots were incubated in secondary antibody solution (goat anti-rabbit horseradish peroxidase (HRP; 1:1000; Vector Laboratories Inc.) for anti-STAT1 and anti-STAT3, goat anti-mouse HRP (1:2000; Santa Cruz Biotechnology Inc.) for anti-STAT1pY701 and anti-STAT3pY705; all diluted in PBS-T) for 1 h at room temperature. .. The blots were washed as described above, incubated in ECL detection solution (Amersham Biosciences, Little Chalfont, Buckinghamshire UK), and exposed to autoradiography film until a suitable image was obtained.

Article Title: Dysregulation of TLR3 Impairs the Innate Immune Response to West Nile Virus in the Elderly ▿
Article Snippet: .. Equal protein concentrations of the supernatants were incubated overnight with 5 μg of anti-STAT1 antibody. .. A fraction of the supernatant was immunoblotted to ensure that equal amounts of STAT1 protein were used in the immunoprecipitation.

Purification:

Article Title: Stat1 Acetylation Inhibits Inducible Nitric Oxide Synthase Expression in Interferon-γ Treated RAW264.7 Murine Macrophages
Article Snippet: .. Purified chromatin was immunoprecipitated using 10 µg of anti-NF-κB p65 (Santa Cruz Biotechnology, Santa Cruz, CA), anti-Stat1 (Santa Cruz Biotechnology, Santa Cruz, CA) or 5 µl of rabbit nonimmune serum; eluted DNA fragments were purified to serve as templates. .. The input fraction corresponded to 0.1 and 0.05% of the chromatin solution before immunoprecipitation.

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 93
    Santa Cruz Biotechnology p stat1
    Co-Immunoprecipitation and ubiquitination of <t>P-STAT1</t> in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.
    P Stat1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p stat1/product/Santa Cruz Biotechnology
    Average 93 stars, based on 44 article reviews
    Price from $9.99 to $1999.99
    p stat1 - by Bioz Stars, 2020-11
    93/100 stars
      Buy from Supplier

    Image Search Results


    Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.

    Journal: Surgery

    Article Title: Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis

    doi: 10.1016/j.surg.2008.03.007

    Figure Lengend Snippet: Co-Immunoprecipitation and ubiquitination of P-STAT1 in CLP Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Blot is representative of four experiments.

    Article Snippet: Using BMM and liver tissue, western blot analysis was performed to determine iNOS, PSTAT1 and OPN expression in CLP in WT and OPN null animals at 0h, 12h and 24h. ( ) WT expression of iNOS and P-STAT1 in BMM and liver is unchanged or decreased from 12h to 24h in association with a 8–10 fold increase in cellular OPN. (p < 0.05; 24h vs. 12h for BMM and liver) In contrast, OPN null expression of iNOS and P-STAT1 is significantly greater in BMM and liver at 24h when compared to that found at 12h. (p < 0.01; 24h vs. 12h for both iNOS and P-STAT1 in BMM and liver) These results show that iNOS and P-STAT1 are significantly increased in the absence of OPN in the CLP model of sepsis.

    Techniques: Immunoprecipitation, Centrifugation, Incubation

    IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.

    Journal: Surgery

    Article Title: Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis

    doi: 10.1016/j.surg.2008.03.007

    Figure Lengend Snippet: IP studies for Ub-Stat1 and iNOS in ex vivo treated BMM from OPN null and WT animals Ex vivo gain-of-function studies were performed using BMM from WT and OPN null mice. MG132 (10 µM), an inhibitor of 26S proteasome function, and/or exogenous OPN (50 µM) were added to assess Ub-P-STAT1 and iNOS expression. BMM cells were treated with LPS (50 ng/ml). The incubation period was 12 hours. Cells were lysed and cleared by centrifugation. Aliquots were precleared by addition of normal IgG and 20 µl of appropriate agarose conjugate. After centrifugation, the supernatant was collected and incubated with 2 µg of P-Stat1 Ab. After washing, immunoblotting was performed using Ub AB. Alternatively, IB was performed for iNOS. Blot is representative of four experiments.

    Article Snippet: Using BMM and liver tissue, western blot analysis was performed to determine iNOS, PSTAT1 and OPN expression in CLP in WT and OPN null animals at 0h, 12h and 24h. ( ) WT expression of iNOS and P-STAT1 in BMM and liver is unchanged or decreased from 12h to 24h in association with a 8–10 fold increase in cellular OPN. (p < 0.05; 24h vs. 12h for BMM and liver) In contrast, OPN null expression of iNOS and P-STAT1 is significantly greater in BMM and liver at 24h when compared to that found at 12h. (p < 0.01; 24h vs. 12h for both iNOS and P-STAT1 in BMM and liver) These results show that iNOS and P-STAT1 are significantly increased in the absence of OPN in the CLP model of sepsis.

    Techniques: Ex Vivo, Mouse Assay, Expressing, Incubation, Centrifugation

    Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.

    Journal: Surgery

    Article Title: Osteopontin Mediates Stat1 Degradation To Inhibit iNOS Transcription In A Cecal Ligation and Puncture Model of Sepsis

    doi: 10.1016/j.surg.2008.03.007

    Figure Lengend Snippet: Western blot of iNOS, phosphorylated STAT1 (P-STAT1) and osteopontin (OPN) expression in CLP in WT and OPN null animals at 0h, 12h and 24h . Blot is presentative of four experiments.

    Article Snippet: Using BMM and liver tissue, western blot analysis was performed to determine iNOS, PSTAT1 and OPN expression in CLP in WT and OPN null animals at 0h, 12h and 24h. ( ) WT expression of iNOS and P-STAT1 in BMM and liver is unchanged or decreased from 12h to 24h in association with a 8–10 fold increase in cellular OPN. (p < 0.05; 24h vs. 12h for BMM and liver) In contrast, OPN null expression of iNOS and P-STAT1 is significantly greater in BMM and liver at 24h when compared to that found at 12h. (p < 0.01; 24h vs. 12h for both iNOS and P-STAT1 in BMM and liver) These results show that iNOS and P-STAT1 are significantly increased in the absence of OPN in the CLP model of sepsis.

    Techniques: Western Blot, Expressing

    ( A ) Expression of p-STAT1 and p-STAT6 in the regenerative tissue was determined by immunohistochemistry. The amplification was 200× . ( B ) Representative western blot images (cropped) of expression of p-STAT1, STAT1, p-STAT6 and STAT6 in the regenerative tissue. ( C ) Gray intensity of p-STAT1 and p-STAT6 was analyzed. *P

    Journal: Scientific Reports

    Article Title: Injectable in situ cross-linking chitosan-hyaluronic acid based hydrogels for abdominal tissue regeneration

    doi: 10.1038/s41598-017-02962-z

    Figure Lengend Snippet: ( A ) Expression of p-STAT1 and p-STAT6 in the regenerative tissue was determined by immunohistochemistry. The amplification was 200× . ( B ) Representative western blot images (cropped) of expression of p-STAT1, STAT1, p-STAT6 and STAT6 in the regenerative tissue. ( C ) Gray intensity of p-STAT1 and p-STAT6 was analyzed. *P

    Article Snippet: Sections for immunohistochemistry were stained with p-STAT1 (Santa Cruz, sc-135648) and p-STAT6 (Santa Cruz, sc-11762).

    Techniques: Expressing, Immunohistochemistry, Amplification, Western Blot

    The relationship between LINC00963 and JAK2/STAT1 pathways in hypoxia‐induced HK‐2 cell model. A, The protein levels of p‐JAK2 and p‐STAT1 in different groups. B, Apoptosis analysed by flow cytometry. C, Expression of apoptosis‐related proteins by Western blot. Data are expressed as mean ± SEM, and different letters represent significant differences ( P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: LINC00963 targeting miR‐128‐3p promotes acute kidney injury process by activating JAK2/STAT1 pathway, et al. LINC00963 targeting miR‐128‐3p promotes acute kidney injury process by activating JAK2/STAT1 pathway

    doi: 10.1111/jcmm.15211

    Figure Lengend Snippet: The relationship between LINC00963 and JAK2/STAT1 pathways in hypoxia‐induced HK‐2 cell model. A, The protein levels of p‐JAK2 and p‐STAT1 in different groups. B, Apoptosis analysed by flow cytometry. C, Expression of apoptosis‐related proteins by Western blot. Data are expressed as mean ± SEM, and different letters represent significant differences ( P

    Article Snippet: As shown in Figure , compared with NC group, the expression of p‐JAK2 and p‐STAT1 significantly increased in renal tissues after transfection with si‐LINC00963.

    Techniques: Flow Cytometry, Expressing, Western Blot

    The relationship between LINC00963 and JAK2/STAT1 pathways on I/R‐induced AKI rat model. A, Expression of p‐JAK2 and p‐STAT1 under different conditions. B, TUNEL staining and cell apoptosis proportion of different groups. C, The protein levels of Bax and Bcl‐2. Data are expressed as mean ± SEM, and different letters represent significant differences ( P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: LINC00963 targeting miR‐128‐3p promotes acute kidney injury process by activating JAK2/STAT1 pathway, et al. LINC00963 targeting miR‐128‐3p promotes acute kidney injury process by activating JAK2/STAT1 pathway

    doi: 10.1111/jcmm.15211

    Figure Lengend Snippet: The relationship between LINC00963 and JAK2/STAT1 pathways on I/R‐induced AKI rat model. A, Expression of p‐JAK2 and p‐STAT1 under different conditions. B, TUNEL staining and cell apoptosis proportion of different groups. C, The protein levels of Bax and Bcl‐2. Data are expressed as mean ± SEM, and different letters represent significant differences ( P

    Article Snippet: As shown in Figure , compared with NC group, the expression of p‐JAK2 and p‐STAT1 significantly increased in renal tissues after transfection with si‐LINC00963.

    Techniques: Expressing, TUNEL Assay, Staining