rabbit anti human sp1  (Cell Signaling Technology Inc)

Journal: Translational Oncology

Article Title: AGEs-RAGE manipulates tumor intrinsic pERK/Sp1/IL6 pathway and reprograms macrophage to promote intrahepatic cholangiocarcinoma progression

doi: 10.1016/j.tranon.2025.102446

Figure Lengend Snippet: AGEs-RAGE axis activates ERK phosphorylation in ICC cells . (A) Stable RAGE knockdown in ICC cells. (B) Western blot analysis of RAGE, p-ERK, E-cadherin, N-cadherin, Vimentin, Sp1 and MMP2 expressions in RBE cells treated with BSA (200 µg/ml) or glucose-AGEs (50,100,200µg/ml) for 24 h. (C) Western blot analysis of p-ERK, E-cadherin, N-cadherin, Vimentin, Sp1 in RBE cells were treated with: BSA (200 µg/ml); glucose-AGEs (200,400µg/ml);or glucose-AGEs(200µg/ml) +U0126(10 µM) for 24 h. (D) RNA-seq analysis of MAPK pathway mRNA expression in shRAGE vs. control cells. ** P < 0.01 compared with control. (E) Western blot analysis of p-ERK, E-cadherin, N-cadherin, Vimentin, Sp1 expressions in shRAGE and control cells. ** P < 0.01 compared with control. (F) Enrichment analysis of downregulated differentially expressed genes (DEGs) in shRAGE vs. control cells.

Article Snippet: Rabbit anti-human RAGE (Abcam, USA) and rabbit anti-human Sp1 (CST, USA) were used as primary antibodies.

Techniques: Phospho-proteomics, Knockdown, Western Blot, RNA Sequencing, Expressing, Control

Journal: Translational Oncology

Article Title: AGEs-RAGE manipulates tumor intrinsic pERK/Sp1/IL6 pathway and reprograms macrophage to promote intrahepatic cholangiocarcinoma progression

doi: 10.1016/j.tranon.2025.102446

Figure Lengend Snippet: S p 1 is a key transcription factor downstream of the AGEs-RAGE axis . (A) Correlation between RAGE and Sp1 gene expression in TCGA data. (B) Representative Sp1 immunohistochemistry in ICC and adjacent liver tissues. (Scale bar, 50µ m). (C) Sp1 mRNA expression (RNA-seq) in shRAGE vs. control cells. ** P < 0.01 compared with control. (D) Western blot analysis of RAGE and Sp1 in RBE cells treated with BSA (200 µg/ml), glucose-AGEs (200µg/ml), both glucose-AGEs(200µg/ml) + RAGE-Ab (5µg/ml) for 24 h. (E) Luciferase activity of IL-6 promoter reporter in RBE cells co-transfected with Sp1 plasmid vs. empty vector for 48 h; * P < 0.05, ** P < 0.01,*** P < 0.001compared with control. (F) IL-6 protein expression (Western blot). RBE cells were transfected with Sp1 gene plasmid or Sp1 siRNA and for 48 h, then treated with: BSA (200µg/ml); glucose-AGEs (200µg/ml); glucose-AGEs (200µg/ml) +RAGE-Ab (5µg/ml) for 24 h.

Article Snippet: Rabbit anti-human RAGE (Abcam, USA) and rabbit anti-human Sp1 (CST, USA) were used as primary antibodies.

Techniques: Gene Expression, Immunohistochemistry, Expressing, RNA Sequencing, Control, Western Blot, Luciferase, Activity Assay, Transfection, Plasmid Preparation

Journal: Translational Oncology

Article Title: AGEs-RAGE manipulates tumor intrinsic pERK/Sp1/IL6 pathway and reprograms macrophage to promote intrahepatic cholangiocarcinoma progression

doi: 10.1016/j.tranon.2025.102446

Figure Lengend Snippet: Glucose-AGEs axis promotes migration and invasion in ICC cells . (A-B) Wound healing assay in RBE cells treated with: BSA (200 µg/mL); glucose-AGEs (200 µg/mL); or glucose-AGEs (200 µg/mL) + RAGE-Ab (5 µg/mL); The wound space was photographed at 0 and 48 h. The wound healing was measured with the following formula: 48-h migration % =(0-h width–48-h width of wound)/(0-h width of wound), *** P < 0.001. (C-D) Transwell migration/invasion assays (RBE cells). Treatments identical to (A-B); All experiments were done in triplicate, and the results are presented as the mean±SD,* P < 0.05, *** P < 0.001. (E) Western blot of RAGE, E-cadherin, N-cadherin, Vimentin, MMP2, and Sp1 in RBE cells. (F-I) RBE cells were subjected to scratch wound-healing assay and transwell assay. RBE cells were transfected with Sp1 siRNA and for 48 h.Then RBE cell were treated with BSA (200µg/ml); glucose-AGEs (200µg/ml), * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Rabbit anti-human RAGE (Abcam, USA) and rabbit anti-human Sp1 (CST, USA) were used as primary antibodies.

Techniques: Migration, Wound Healing Assay, Western Blot, Transwell Assay, Transfection

Journal: Translational Oncology

Article Title: AGEs-RAGE manipulates tumor intrinsic pERK/Sp1/IL6 pathway and reprograms macrophage to promote intrahepatic cholangiocarcinoma progression

doi: 10.1016/j.tranon.2025.102446

Figure Lengend Snippet: RAGE overexpression correlates with poor ICC prognosis . (A) RAGE mRNA expression in ICC (TCGA). (B) Kaplan-Meier survival analysis by RAGE expression (high vs. low). (C) Representative RAGE IHC in ICC vs. adjacent liver (Scale bar: 50 µm). (D) RAGE staining intensity distribution in 153 ICC patients. (E) Confocal imaging showing RAGE expression in normal bile duct vs. ICC tissues (Scale bar: 50 µm). Concurrent upregulation of RAGE and Sp1 in ICC tissues (Western blot).

Article Snippet: Rabbit anti-human RAGE (Abcam, USA) and rabbit anti-human Sp1 (CST, USA) were used as primary antibodies.

Techniques: Over Expression, Expressing, Staining, Imaging, Western Blot