p stat3 tyr705 antibody  (Cell Signaling Technology Inc)

Journal: Signal Transduction and Targeted Therapy

Article Title: Bioengineered iPSC-derived human macrophages with increased angiotensin-converting enzyme (ACE) expression suppress solid tumor growth

doi: 10.1038/s41392-026-02650-3

Figure Lengend Snippet: Determination of M1 vs. M2 polarization in iMac. a Pro-inflammatory M1 cytokines IL-1β, TNFα, IL-6, and IL-8 measured by ELISA. b Anti-inflammatory M2 cytokines IL-4, IL-10, IL-13, CCL5, CCL17, and CCL20 measured by ELISA. For a , b iMac were cultured in 6-well plates (3 × 10 6 cells/well). To challenge with tumor factors, iMac were co-cultured with 50% fresh supernatant from SK-MEL-28 cells every 24 h for 3 days. Supernatants were collected during the final 24 h for cytokine measurement. c Activation (phosphorylation) of NF-kB p65, STAT1, STAT3, and STAT6 in iMac was determined using Western blotting after conditioning with melanoma (SK-MEL-28) supernatant for 72 h. Protein band intensity was quantified using Odyssey version 3.0 software. After adjusting for loading using the β-actin band, data are presented as fold change to the iMac (Dox-) group, which is set as 1. For a – c , one-way ANOVA with Bonferroni’s correction for multiple comparisons was used to analyze group comparisons. Data are presented as means ± SD. Each experiment was performed three times, with triplicate samples each time. * p < 0.05, *** p < 0.001, and **** p < 0.0001

Article Snippet: The polyvinylidene difluoride (PVDF) membranes were incubated with specific primary antibodies against ACE (R&D Systems, MAB9291, 1:1,000), GAPDH (Sigma-Aldrich, SAB5600208, 1:2,000), β-actin (Sigma-Aldrich, A3854; 1:1000), phosphorylated NF-kB p65 (Novus, NB100-82086, 1:500), phosphorylated STAT1 (R&D Systems, AF2894, 1 μg/ml), phosphorylated STAT3 (Novus, NBP2-24463, 0.5 μg/ml), or phosphorylated STAT6 (Millipore, 06-937, 1:1000).

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Activation Assay, Phospho-proteomics, Western Blot, Software

Journal: Advances in Radiation Oncology

Article Title: Kupffer Cell-Derived Interleukin-6 Aggravates Radiation-Induced Liver Disease by Activating Hepatocyte STAT3 to Promote Ccng1 Transcription

doi: 10.1016/j.adro.2026.102003

Figure Lengend Snippet: Role of the JAK–STAT signaling pathway in hepatocytes during radiation-induced liver disease (RILD). (A) Volcano plot of differential gene expression analysis in hepatocytes after irradiation (IR). Red and blue indicate upregulated and downregulated genes, respectively. The dotted horizontal line represents a P value of 0.05. The dotted vertical lines represent a log2-fold change of 1.5 or −1.5. (Right) Bubble chart depicting the top 5 Kyoto Encyclopedia of Genes and Genomes (KEGG)-enriched pathways corresponding to down/upregulated genes in hepatocytes after IR (screening the pathways and sorting them from large to small according to the −log 10 P value). (B) Western blot analysis showing p-STAT3 and GAPDH expression in primary hepatocytes isolated from Ctrl rats or IR rats treated with placebo, anti-interleukin (IL)-6, or sgp130Fc (n = 5). (C) Western blot analysis showing p-STAT3 and GAPDH expression in primary hepatocytes isolated from IR rats treated with placebo, ruxolitinib (RUX), or tofacitinib (TOF) (n = 5). (D) Quantitative enzyme-linked immunosorbent assay (ELISA) analysis of serum liver enzymes in IR rats treated with placebo, RUX, or TOF (n = 5). (E) Hematoxylin–eosin (H&E) staining of the livers of IR rats treated with placebo, RUX, or TOF (n = 5). (F) Costaining of HNF4α (red) and TUNEL (green) in the livers of IR rats treated with placebo, RUX, or TOF (n = 5). (G) Overlap of p-STAT3-binding genes identified using ChIP-Seq with upregulated genes ( P < .05 and log2-fold change > 2) in hepatocytes identified using scRNA-seq. (H) Violin plots showing the expression levels of overlapping genes identified using scRNA-seq. (I) qRT‒PCR analysis of overlapping genes in isolated primary hepatocytes from IR rats treated with placebo, RUX, or TOF (n = 5). Analysis of variance (ANOVA); *, P < .05; ⁎⁎ , P < .01; ⁎⁎⁎ , P < .001; ns, nonsignificant.

Article Snippet: After sonication, immunoprecipitation was performed with a p-STAT3 (Tyr705) antibody (Cell Signaling Technology, #9145).

Techniques: Gene Expression, Irradiation, Western Blot, Expressing, Isolation, Enzyme-linked Immunosorbent Assay, Staining, TUNEL Assay, Binding Assay, ChIP-sequencing

Journal: Advances in Radiation Oncology

Article Title: Kupffer Cell-Derived Interleukin-6 Aggravates Radiation-Induced Liver Disease by Activating Hepatocyte STAT3 to Promote Ccng1 Transcription

doi: 10.1016/j.adro.2026.102003

Figure Lengend Snippet: Schematic of the Kuppfer cell (KC)-hepatocyte crosstalk mechanism in RILD. Irradiation (IR) stimulates KCs to secrete IL-6, which binds to the IL-6R/gp130 complex on hepatocytes to activate JAK; phosphorylated JAK induces STAT3 phosphorylation, and nuclear-translocated p-STAT3 binds to the Ccng1 promoter to promote its transcription; CCNG1 then regulates MDM2 to mediate ubiquitination-dependent TP53 proteolysis, ultimately enhancing hepatocyte apoptosis and driving radiation-induced liver disease (RILD) progression.

Article Snippet: After sonication, immunoprecipitation was performed with a p-STAT3 (Tyr705) antibody (Cell Signaling Technology, #9145).

Techniques: Irradiation, Phospho-proteomics, Ubiquitin Proteomics

Journal: Scientific Reports

Article Title: Adapalene, an RAR agonist, exerts anti-inflammatory effects by regulating macrophage polarization through RAR -mediated signaling pathways

doi: 10.1038/s41598-026-44454-z

Figure Lengend Snippet: Adapalene reduces LPS-stimulated MAPK and PI3K/Akt signaling in murine macrophages. RAW264.7 cells were pretreated with or without adapalene (10, 100, or 1000 nM) for 1 h and then stimulated with LPS (100 ng/mL) for 30 min. Total cellular proteins were resolved using SDS-PAGE and detected using specific antibodies against p-p38, p38, p-JNK, JNK, p-ERK, ERK (A), p-Akt, Akt, p-PI3K, PI3K, p-STAT3, and STAT3 (B). β-actin was used as an internal control. Membranes were cut prior to antibody incubation; all available uncropped images are provided in the Supplementary Information. Original full-length images for certain replicates are unavailable due to loss of original acquisition files. Representative results are shown and density analysis is presented as the mean ± S.D. of three independent experiments in lower panel. AD; adapalene, * P < 0.05, ** P < 0.01, and *** P < 0.001 vs. control; # P < 0.05 and ## P < 0.01 vs. LPS alone.

Article Snippet: p-STAT3 , Cell Signaling , 9131 , Rabbit , 1:1000.

Techniques: SDS Page, Control, Incubation

Journal: Scientific Reports

Article Title: Adapalene, an RAR agonist, exerts anti-inflammatory effects by regulating macrophage polarization through RAR -mediated signaling pathways

doi: 10.1038/s41598-026-44454-z

Figure Lengend Snippet: RARβ mediates adapalene-induced anti-inflammatory signaling. (A, B) RAW264.7 cells were transfected with control siRNA or RARβ siRNA (100 nM), and then treated with LPS in the presence or absence of adapalene for 24 h. Total cellular proteins were resolved using SDS-PAGE and detected using the specific antibodies p-p38, p38, p-JNK, JNK, p-ERK, and ERK (A) and specific antibodies p-Akt, Akt, p-PI3K, PI3K, p-STAT3, and STAT3 (B). (C, D) RAW264.7 cells were treated in the presence or absence of LE135 (0.1–10 µM) with or without adapalene and LPS for 24 h. Total cellular proteins were resolved using SDS-PAGE and detected using specific antibodies p-p38, p38, p-JNK, JNK, p-ERK, and ERK (C) and specific antibodies p-Akt, Akt, p-PI3K, PI3K, p-STAT3, and STAT3 (D). Membranes were cut prior to antibody incubation; all available uncropped images are provided in the Supplementary Information. Original full-length images for certain replicates are unavailable due to loss of original acquisition files. Density analyses are shown in lower panel. The experiment was performed three times in triplicate and results are presented as the mean ± S.D. of three independent experiments. AD; adapalene, * P < 0.05, ** P < 0.01, and *** P < 0.001 vs. control; # P < 0.05, ## P < 0.01, and ### P < 0.001 vs. LPS alone; & P < 0.05, && P < 0.01, and &&& P < 0.001 vs. AD plus LPS.

Article Snippet: p-STAT3 , Cell Signaling , 9131 , Rabbit , 1:1000.

Techniques: Transfection, Control, SDS Page, Incubation

Journal: Scientific Reports

Article Title: Adapalene, an RAR agonist, exerts anti-inflammatory effects by regulating macrophage polarization through RAR -mediated signaling pathways

doi: 10.1038/s41598-026-44454-z

Figure Lengend Snippet: In vivo anti-inflammatory effects of adapalene in LPS-induced septic shock model. Six mice per group were treated with vehicle only or adapalene (20, 50, or 100 mg/kg, p.o. ) for 1 h and then injected with LPS (25 mg/kg, i.p. ). Serum and liver samples were collected from each mouse after 6 h. (A, B) TNFα, IL-6, and IL-1β protein in serum (A) and mRNA levels of TNFα, IL-1β, IL-6, iNOS, and COX-2 in liver (B) were determined using EIA and qRT-PCR, respectively. (C) TNFα, IL-1β, IL-6, iNOS, COX-2, MRC1, Arg1, RARα, RARβ, and RARγ protein levels in liver were determined using western blot. Membranes were cut prior to antibody incubation; all available uncropped images are provided in the Supplementary Information. Original full-length images for certain replicates are unavailable due to loss of original acquisition files. Density analysis is shown in the right panel. (D) Protein levels of p38, p-p38 MAPK, ERK, p-ERK, JNK, p-JNK, PI3K, p-PI3K, Akt, p-Akt, STAT3, and p-STAT3 in the liver were determined using western blotting. Density analysis is shown in the lower panel. Results are presented as the means ± S.D. of six mice. Representative images of p-NF-κB staining (E), F4/80 staining (F), H&E staining (H), and Picro-Sirius Red staining (K) of liver tissues were shown. Quantitative analysis of p-NF-κB and F4/80 staining was performed using ImageJ software (G). The survival rate after adapalene administration in LPS-induced sepsis is shown (I). Hepatic mRNA and protein levels of fibrosis markers were determined (J). AD; adapalene, * P < 0.05, ** P < 0.01, and *** P < 0.001 vs. control; # P < 0.05, ## P < 0.01, and ### P < 0.001 vs. LPS alone.

Article Snippet: p-STAT3 , Cell Signaling , 9131 , Rabbit , 1:1000.

Techniques: In Vivo, Injection, Quantitative RT-PCR, Western Blot, Incubation, Staining, Software, Control

Journal: Scientific Reports

Article Title: Adapalene, an RAR agonist, exerts anti-inflammatory effects by regulating macrophage polarization through RAR -mediated signaling pathways

doi: 10.1038/s41598-026-44454-z

Figure Lengend Snippet: In vivo anti-inflammatory effects of adapalene in HFD-induced obesity model. Adapalene (10 or 50 mg/kg) was orally administered once daily to HFD-induced obese mice ( n = 10 per group) for 3 weeks. On the last day of administration, serum and liver samples were collected from each mouse. (A) TNFα, IL-1β, IL-6, iNOS, COX-2, MRC1, Arg1, p38, p-p38 MAPK, ERK, p-ERK, JNK, p-JNK, RARα, RARβ, and RARγ protein levels in the liver were determined using western blotting. Density analysis is shown in the right panel. (B) PI3K, p-PI3K, Akt, p-Akt, STAT3, and p-STAT3 protein levels in the liver were determined using western blotting. Membranes were cut prior to antibody incubation; all available uncropped images are provided in the Supplementary Information. Original full-length images for certain replicates are unavailable due to loss of original acquisition files. Density analysis is shown in the lower panel. (C) TNFα, IL-6, IL-1β, COX-2, iNOS, MRC1, and Arg1 mRNA levels in the liver were determined using qRT-PCR. (D) Representative images of Oil Red O staining and Picro-Sirius Red staining (G) of liver tissues are shown. Results are presented as the means ± S.D. of eight mice. (E, F) Hepatic protein levels (E), mRNA levels of fibrosis markers (F) and serum ALT and AST levels (F) were determined. AD; adapalene, * P < 0.05, ** P < 0.01, and *** P < 0.001 vs. control; # P < 0.05, ## P < 0.01, and ### P < 0.001 vs. LPS alone.

Article Snippet: p-STAT3 , Cell Signaling , 9131 , Rabbit , 1:1000.

Techniques: In Vivo, Western Blot, Incubation, Quantitative RT-PCR, Staining, Control

Journal: Scientific Reports

Article Title: Adapalene, an RAR agonist, exerts anti-inflammatory effects by regulating macrophage polarization through RAR -mediated signaling pathways

doi: 10.1038/s41598-026-44454-z

Figure Lengend Snippet: Proposed working model of the mechanism of action of adapalene. Adapalene produces anti-inflammatory effects through dual mechanisms involving the suppression of the M1-mediated inflammatory response and induction of the M2-mediated anti-inflammatory response. Both actions are mediated through RARβ activation, followed by MAPK and PI3K/Akt inactivation, and NF-κB inactivation (M1 response) as well as by STAT3 phosphorylation (M2 response).

Article Snippet: p-STAT3 , Cell Signaling , 9131 , Rabbit , 1:1000.

Techniques: Activation Assay, Phospho-proteomics