p ddr1 2  (R&D Systems)

Journal: Cancer Cell International

Article Title: PHI-501, a dual inhibitor of RAF and DDR1/2, overcomes MAPK drug resistance in Melanoma

doi: 10.1186/s12935-026-04271-w

Figure Lengend Snippet: MAPK inhibitor-resistant melanoma cells exhibit enhanced DDR1/2 signaling. ( A ) Clonogenic assay showing the validation of drug-resistant cell lines. ( B ) Quantification of clonogenic assay shown in Fig. 1A. Data are presented as relative colony formation (%) normalized to the control group and expressed as the mean ± SEM, n = 3 replicates. Statistical significance was determined by two-way ANOVA. * p < 0.05, # p < 0.001, † p < 0.001, ‡ p < 0.0001. ( C ) Cell viability assay of RAF and MEK inhibitors in parental cells and drug-resistant cells for 48 h. Data are presented as the mean ± SEM, n = 3 replicates. ( D ) Western blot analysis of pERK and pAKT signaling responses in parental and resistant melanoma cells following 48-h treatment with vehicle or the corresponding resistance-selecting drug(s) at 1 µM. ( E ) Bubble plot representing Gene Set Enrichment Analysis (GSEA) using hallmark gene set comparing parental and resistant melanoma cells. Pathway enrichment was calculated using GSEA, with color indicating NES (blue = downregulated, red = upregulated) and bubble size representing −log10(p-value). ( F ) SK-MEL-2 and SK-MEL-2BR cells were stimulated with type I collagen (40 µg/ml) for 18 h to activate DDR signaling

Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies diluted in 5% BSA in TBST as indicated in the text: p-DDR1 (Cell Signaling Technology, CST14531 ), p-DDR1/2 (R&D Systems, MAB25382), p-AKT (Cell Signaling Technology, CST4058), AKT (Cell Signaling Technology, CST9272), p-MEK (Cell Signaling Technology, CST9121), p-ERK (Cell Signaling Technology, CST9101), ERK (Cell Signaling Technology, CST9102), Cyclin D1(Cell Signaling Technology, CST2978), and Survivin (Cell Signaling Technology, CST2808).

Techniques: Clonogenic Assay, Biomarker Discovery, Control, Viability Assay, Western Blot

Journal: Cancer Cell International

Article Title: PHI-501, a dual inhibitor of RAF and DDR1/2, overcomes MAPK drug resistance in Melanoma

doi: 10.1186/s12935-026-04271-w

Figure Lengend Snippet: DDR1 and DDR2 are associated with MAPK pathway activation and therapeutic resistance in melanoma. ( A ) Kaplan–Meier survival analysis showing patient outcomes based on DDR1 and DDR2 expression levels across all TCGA cancer types. Patients were stratified into high (top 25%) and low (bottom 25%) expression groups, and overall survival (OS) was analyzed. ( B ) Proteomic analysis of BRAF/NRAS-mutant SKCM. Box plots demonstrate differential expression of AKT and phosphorylated MEK1 (pMEK1-S217/S221) between wild-type and mutant groups. ( C ) Correlation heatmaps between DDR1/2 and key genes in the MAPK and AKT signaling pathways in TCGA-SKCM samples. Color intensity represents the correlation value, with red indicating a positive correlation. Significance levels are indicated by asterisks (* p < 0.05, ** p < 0.01, *** p < 0.001). ( D ) Correlation analysis revealing DDR1 expression (DepMap) and MAPK pathway inhibitor sensitivity (GDSC2) in melanoma cell lines. Scatter plots depicting correlations between DDR1 expression levels and IC50 to Dabrafenib (left) and Trametinib (right)

Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies diluted in 5% BSA in TBST as indicated in the text: p-DDR1 (Cell Signaling Technology, CST14531 ), p-DDR1/2 (R&D Systems, MAB25382), p-AKT (Cell Signaling Technology, CST4058), AKT (Cell Signaling Technology, CST9272), p-MEK (Cell Signaling Technology, CST9121), p-ERK (Cell Signaling Technology, CST9101), ERK (Cell Signaling Technology, CST9102), Cyclin D1(Cell Signaling Technology, CST2978), and Survivin (Cell Signaling Technology, CST2808).

Techniques: Activation Assay, Expressing, Mutagenesis, Quantitative Proteomics, Protein-Protein interactions

Journal: Discover Oncology

Article Title: Single-cell and immune-context integration identifies basement-membrane/metastasis signatures that sharpen bladder-cancer diagnosis and prognosis

doi: 10.1007/s12672-026-04440-3

Figure Lengend Snippet: Immune characteristics of BLCA patients stratified by risk scores. A Differential immune cell infiltration between risk groups. B Comparative analysis of immune pathway activities in high- and low-risk groups. C Correlation analysis between risk scores and immune subtypes. D Associations between SERPINF1 expression and multiple immune cells. E Scatterplot showing correlation between SERPINF1 expression and M2 macrophage infiltration. F Scatterplot showing correlation between SERPINF1 expression and activated dendritic cell infiltration

Article Snippet: The primary antibodies used were DDR2 (Proteintech, 67126-1-Ig), SERPINF1 (Proteintech, 26045-1-AP), CD68 (Abcam, ab213363), and α-SMA (Abcam, ab124964).

Techniques: Expressing

Journal: Discover Oncology

Article Title: Single-cell and immune-context integration identifies basement-membrane/metastasis signatures that sharpen bladder-cancer diagnosis and prognosis

doi: 10.1007/s12672-026-04440-3

Figure Lengend Snippet: Cell lineage and spatial distribution of DDR2 and SERPINF1 in BLCA. A UMAP plot of single-cell transcriptomes colored by cell type, highlighting major populations including epithelial cells, fibroblasts, endothelial cells, monocytes, B cells, and adipocytes. Both DDR2 and SERPINF1 were predominantly expressed in fibroblasts. B , C Spatial transcriptomics analysis. Left: cell type annotation map of normal tissue; right: spatial expression heatmaps of DDR2 and SERPINF1 in tumor tissue. D , E Representative multiplex immunofluorescence (Opal-TSA) images acquired from the same section and the same field (ROI), showing DAPI nuclear staining and α-SMA and CD68 signals, with DDR2 (D) or SERPINF1 (E) channels displayed separately for clarity; merged images support the spatial transcriptomic findings. Scale bar, 50 μm

Article Snippet: The primary antibodies used were DDR2 (Proteintech, 67126-1-Ig), SERPINF1 (Proteintech, 26045-1-AP), CD68 (Abcam, ab213363), and α-SMA (Abcam, ab124964).

Techniques: Single Cell, Spatial Transcriptomics, Expressing, Multiplex Assay, Immunofluorescence, Staining

Journal: Cell Reports Medicine

Article Title: Drug screening in 3D microtumors reveals DDR1/2-MAPK12-GLI1 as a vulnerability in cancer-associated fibroblasts

doi: 10.1016/j.xcrm.2025.102357

Figure Lengend Snippet: Doramapimod unexpectedly targets DDR1/2 and MAPK12, regulating extracellular matrix gene expression in cancer-associated fibroblasts (A) Kinome profiling of doramapimod . Left: residual kinase activity for 370 kinases treated with 500 nM doramapimod using a radioactive ATP assay. Kinases with <20% residual activity are indicated in red. Right: bar chart highlighting top inhibited kinases. (B) CAF gene expression after kinase knockdown . Heatmap showing changes in ACTA2 and CXCL12 expression in breast cancer-derived CAFs following siRNA knockdown of doramapimod target kinases. (C) Plot showing qPCR analysis of CXCL12 expression in breast CAFs following siRNA-mediated knockdown of DDR1, DDR2, and MAPK12, with or without doramapimod treatment. Data represent mean ± SEM; n = 3 per group. Student’s t test, compared with DMSO control. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. (D) Transcriptional changes in CAFs upon DDR1 , DDR2 , or MAPK12 depletion . Volcano plots showing DEGs (>2-fold, p < 0.05) after siRNA knockdown of DDR1 (left), DDR2 (middle), or MAPK12 (right) in primary breast CAFs. Downregulated genes are shown in blue and upregulated in red. n = 3 per group. (E) Pathway enrichment of downregulated genes following combined DDR1/2 and MAPK12 knockdown . Bar graph showing enrichment across Gene Ontology, Reactome, and KEGG pathways, with ECM-related processes highlighted in red. (F) Neutralization of CAFs’ growth stimulatory effect through depletion of DDR1/2 and MAPK12 kinase expression . Top: schematic of the experimental design showing breast CAFs treated with siRNAs against the indicated kinases, then co-cultured with 4T1 cancer cells labeled with nuclear GFP. Bottom: growth curve of 4T1 cells co-cultured with kinase depleted CAFs, displayed as Mean ± SEM. n = >3 in each group. (G) DDR1/2 enhances p38 phosphorylation . Western blot analysis showing elevated levels of phospho-p38 in CAFs overexpressing DDR1 or DDR2 compared to GFP control. Total p38 and β-actin are shown as loading controls. (H) Proposed model of doramapimod action . Doramapimod inhibits the DDR1/2–MAPK12 signaling axis, which drives ECM production in CAFs.

Article Snippet: Rabbit monoclonal DDR2 (E6V2N) Antibody , Cell Signaling Technology , Cat#25814;RRID:AB_3674514.

Techniques: Gene Expression, Activity Assay, ATP Assay, Knockdown, Expressing, Derivative Assay, Control, Neutralization, Cell Culture, Labeling, Phospho-proteomics, Western Blot

Journal: Cell Reports Medicine

Article Title: Drug screening in 3D microtumors reveals DDR1/2-MAPK12-GLI1 as a vulnerability in cancer-associated fibroblasts

doi: 10.1016/j.xcrm.2025.102357

Figure Lengend Snippet: DDR1/2 and MAPK12 converge at GLI to regulate ECM production and support cancer cell growth (A) Gene overlap and expression analysis . Left: a Venn diagram showing the overlap of genes downregulated in response to knockdown of DDR1/2 and MAPK12 in CAFs. Right: a heatmap displaying changes in the expression of 7 genes commonly downregulated by depletion of DDR1/2 and MAPK12. The transcriptional factors, ECM proteins, and immune regulatory roles of these genes are highlighted. (B) Doramapimod reduces GLI1 nuclear localization in CAFs . Left: representative immunofluorescence images showing reduced nuclear GLI1 signal in CAFs following doramapimod treatment (1 μM for 48 h). Right: quantification of nuclear GLI1 intensity from at least 20 cells per group, indicating significantly reduced nuclear localization. Scale bars, 20 μm; ∗∗ p < 0.01; Student’s t test. (C) GLI1 transcriptional activity is inhibited by doramapimod , DDR1/2 , and MAPK12 knockdown . Left: luciferase reporter assay in pancreatic CAFs shows decreased GLI1 transcriptional activity upon treatment with doramapimod (1 μM) or GANT61 (1 μM). Right: similar reduction in GLI1 activity observed upon knockdown of DDR1, DDR2, or MAPK12. Data represent mean ± SEM from two-three biological replicates; ∗ p < 0.05 and ∗∗ p < 0.01; unpaired Student’s t test. (D) GLI1 inhibition downregulates ECM-associated pathways . Pathway enrichment plots based on RNA-seq of breast and pancreatic CAFs treated with GANT61. Genes involved in ECM regulation, including integrin signaling, collagen formation, and matrix remodeling, are significantly downregulated (adjusted p values indicated by color scale). (E) DDR1/2 promotes nuclear localization of GLI . Representative images showing that overexpression of DDR1/2 in normal human pancreatic fibroblasts promotes nuclear localization of phosphorylated p38 MAPK and GLI1. Scale bars, 20 μm. (F) Neutralization of CAFs' growth stimulatory effect by GLI depletion . Left: plot showing growth of GFP-labeled 4T1 cancer cells on CAFs with intact or depleted levels of GLI1, displayed as mean ± SEM. N = >3 in each group. Student’s t tests with Holm-Sidak correction. ∗∗ p < 0.01. Right: Gant61 treatment does not affect 4T1 cancer cell growth directly in serum-supported monoculture, but significantly reduces 4T1 cell growth when co-cultured with CAFs in serum-free condition, displayed as mean ± SEM. n indicates at least 3 replicates in each group. (G) M odel of the non-canonical hedgehog pathway in CAFs . Schematic illustrating DDR1/2-mediated activation of p38/MAPK12 and GLI drives ECM production in CAFs and supports cancer cell growth.

Article Snippet: Rabbit monoclonal DDR2 (E6V2N) Antibody , Cell Signaling Technology , Cat#25814;RRID:AB_3674514.

Techniques: Expressing, Knockdown, Immunofluorescence, Activity Assay, Luciferase, Reporter Assay, Inhibition, RNA Sequencing, Over Expression, Neutralization, Labeling, Cell Culture, Activation Assay