primary antibodies against stat3  (Proteintech)

Journal: Experimental and Therapeutic Medicine

Article Title: Yuanhuacine modulates lipopolysaccharide-induced interleukin-6 through regulation of the JAK1/STAT3 pathway and prevents tubular damage in acute kidney injury

doi: 10.3892/etm.2025.12918

Figure Lengend Snippet: YC inhibits LPS-induced phosphorylation of JAK1 and STAT3 in THP-1 cells. (A) Differentiated THP-1 cells were treated with 5 ng/ml LPS and 200 nM YC for up to 12 h. The activation of kinases as well as transcription factors was evaluated by western blotting analysis following LPS stimulation. (B) Quantification of phosphorylated JAK1 and STAT3 proteins after treatment with increasing concentrations of YC. White lines indicate separate groups. Phosphorylation levels were normalized to total JAK1 and STAT3 expression and quantified using ImageJ software. Data are presented as the mean ± standard deviation from three independent experiments. Statistical significance was determined using one-way analysis of variance. ### P<0.001, ## P<0.01 vs. control; $$ P<0.01 vs. LPS; ns, not significant; YC, yuanhuacine; LPS, lipopolysaccharide; JAK1, Janus kinase 1; STAT3, signal transducer and activator of transcription 3; p-, phosphorylated.

Article Snippet: Fixed cells were incubated overnight at 4 ̊C with primary antibodies against p-STAT3 and STAT3 (both at 1:200 dilution; Cell Signaling Technology).

Techniques: Phospho-proteomics, Activation Assay, Western Blot, Expressing, Software, Standard Deviation, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Yuanhuacine modulates lipopolysaccharide-induced interleukin-6 through regulation of the JAK1/STAT3 pathway and prevents tubular damage in acute kidney injury

doi: 10.3892/etm.2025.12918

Figure Lengend Snippet: YC attenuates LPS-induced cytoplasmic-to-nuclear translocation of phosphorylated STAT3 in THP-1 cells. THP-1 cells were treated with 5 ng/ml LPS for 2 h in the absence or presence of YC at three concentrations: 8, 40, and 200 nM. The subcellular localization of STAT3 was assessed by multiple methods. (A) Western blot analysis of cytoplasmic and nuclear fractions was performed to determine the localization of STAT3 protein. (B) Confocal microscopy image showing the intracellular distribution of p-STAT3. (C) Confocal microscopy image showing the intracellular distribution of total STAT3. β-Actin was used as a cytoplasmic marker, and Lamin B served as a nuclear marker. Nuclei were stained with DAPI (blue), and STAT3 (total and p-STAT3) was visualized in green. Scale bars: 10 µm. YC, yuanhuacine; LPS, lipopolysaccharide; STAT3, signal transducer and activator of transcription 3; p-, phosphorylated.

Article Snippet: Fixed cells were incubated overnight at 4 ̊C with primary antibodies against p-STAT3 and STAT3 (both at 1:200 dilution; Cell Signaling Technology).

Techniques: Translocation Assay, Western Blot, Confocal Microscopy, Marker, Staining

Journal: Experimental and Therapeutic Medicine

Article Title: Yuanhuacine modulates lipopolysaccharide-induced interleukin-6 through regulation of the JAK1/STAT3 pathway and prevents tubular damage in acute kidney injury

doi: 10.3892/etm.2025.12918

Figure Lengend Snippet: Protective effects of YC in an LPS-induced AKI mouse model. Renal dysfunction was evaluated by measuring (A) BUN and (B) sCr levels, and (C) tubular injury was assessed by scoring histological damage (n=5). (D) A representative image of H&E staining and immunohistochemistry images in kidney tissue. The arrows indicate tubular degeneration. In kidney tissues of mice with AKI (n=5), the expression levels of (E) KIM-1, (F) NGAL and (G) IL-6, markers of renal injury, were analyzed by quantitative polymerase chain reaction. (H) Western blot analysis of JAK1 and STAT3 activation in kidney tissues of AKI mice (n=4) following YC treatment. The image density was analyzed using ImageJ. Scale bars: 500 µm (40x), 100 µm (100x, 200x), and 50 µm (400x). Data are presented as the mean ± standard deviation. Statistical significance was determined using one-way analysis of variance. ### P<0.001, ## P<0.01, # P<0.05 vs. control; $$$ P<0.001, $$ P<0.01, $ P<0.05 vs. LPS; ns, not significant; AKI, acute kidney injury; H&E, hematoxylin and eosin; KIM-1, kidney injury molecule 1; NGAL, neutrophil gelatinase-associated lipocalin; BUN, blood urea nitrogen; sCr, serum creatinine; YC, yuanhuacine; LPS, lipopolysaccharide; IL-6, interleukin-6; JAK1, Janus kinase 1; STAT3, signal transducer and activator of transcription 3.

Article Snippet: Fixed cells were incubated overnight at 4 ̊C with primary antibodies against p-STAT3 and STAT3 (both at 1:200 dilution; Cell Signaling Technology).

Techniques: Staining, Immunohistochemistry, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Activation Assay, Standard Deviation, Control