Journal: Life

Article Title: Investigation of Cell Mechanics and Migration on DDR2-Expressing Neuroblastoma Cell Line

doi: 10.3390/life14101260

Figure Lengend Snippet: shRNA-mediated knockdown of DDR2. ( A ) DDR2 mRNA expression across cancer lines was ana-lyzed in the Dependency Map Portal ( https://depmap.org/portal ; Broad Institute, accessed on 15 February 2024). ( B ) Western blot of DDR2 expression (vs. GAPDH) upon 4 days of doxycycline treatment (0–12.5 mg/mL) on Tet-shDDR2 SHSY5Y cells. ( C ) Cell survival/proliferation time course assay (as measured by % confluency) of Tet-shDDR2 SHSY5Y cells treated with doxycycline (on day 0) at increasing concentrations as indicated ( n = 6 wells/condition). ( D ) Western blot of DDR2 and phospho-tyrosine 740-DDR2 (phY740-DDR2) from Tet-shDDR2 SHSY5Y cells pretreated for 3 days with the indicated concentrations of doxycycline and left unstimulated or induced with 25 mg/mL of rat-tail type I collagen for 16 h. Protein quantification data for Western blots can be found in .

Article Snippet: Primary antibodies were: from Cell Signaling Technology DDR2 (#12133); from MilliporeSigma (Darmstadt, Germany) Anti-GAPDH antibody (MAB374); and from R & D Systems human phopho-DDR1/DDR2 (Y796/Y740) antibody (MAB25382).

Techniques: shRNA, Knockdown, Expressing, Western Blot

Journal: Life

Article Title: Investigation of Cell Mechanics and Migration on DDR2-Expressing Neuroblastoma Cell Line

doi: 10.3390/life14101260

Figure Lengend Snippet: DDR2 downregulation decreases neuroblastoma cell stiffness. Young’s modulus measurements of ( A ) shCTRL and shDDR2 cells using atomic force microscopy. Indentation of ( B ) cell front (red arrow), ( C ) cell middle (blue arrow), and ( D ) cell back (yellow arrow). Young’s modulus of experiments were performed in three independent experiments. Unpaired t -test, * p < 0.05; *** p < 0.001 ( n = 32–33 cells). Data are presented as ± s.e.m.

Article Snippet: Primary antibodies were: from Cell Signaling Technology DDR2 (#12133); from MilliporeSigma (Darmstadt, Germany) Anti-GAPDH antibody (MAB374); and from R & D Systems human phopho-DDR1/DDR2 (Y796/Y740) antibody (MAB25382).

Techniques: Microscopy

Journal: Frontiers in Oncology

Article Title: COL10A1-DDR2 axis promotes the progression of pancreatic cancer by regulating MEK/ERK signal transduction

doi: 10.3389/fonc.2022.1049345

Figure Lengend Snippet: COL10A1 regulates DDR2 expression in PDAC cells. (A) Western blot results showed the effect of COL10A1 on DDR2 and P-DDR2 protein levels. (B) IHC staining of DDR2 in PDAC and normal pancreatic tissues. The scale bar at 200× magnification represents 20 µm. The scale bar at 400× magnification represents 50 µm. (C) DDR2 IHC scores were differentially expressed between the normal and tumor groups. n = 30. Two-tailed Student’s t-test. (D) Pearson correlation analysis was conducted to analyze the relation between COL10A1 and DDR2 in 30 PDAC tissues. (E) Co-localization of COL10A1 and DDR2 in PDAC cells detected by immunofluorescence. (F, G) The detection of the interaction between COL10A1 and DDR2 by Co-IP. *** P <0.001.

Article Snippet: The following primary antibodies were used: DDR2, 1:200 (ABclonal); COL10A1, 1:200 (BOSTER); E-cadherin, N-cadherin, Vimentin, 1:200 (Cell Signaling Technology).

Techniques: Expressing, Western Blot, Immunohistochemistry, Two Tailed Test, Immunofluorescence, Co-Immunoprecipitation Assay

Journal: Frontiers in Oncology

Article Title: COL10A1-DDR2 axis promotes the progression of pancreatic cancer by regulating MEK/ERK signal transduction

doi: 10.3389/fonc.2022.1049345

Figure Lengend Snippet: COL10A1 facilitates the malignant biological behavior of PDAC cells through DDR2. (A) Effect of DDR2 overexpression on the proliferative capacity of SW1990, BXPC cells after disruption of COL10A1 using clone formation assay. (B) Effects of shCOL and shCOL10A1+DDR2 on the proliferation ability of SW1990 and BXPC-3 cells by CCK8 assay. (C-F) Effects of shCOL and shCOL10A1+DDR2 on the migration ability of SW1990 and BXPC-3 cells by wound healing assays. Scale bar, 50 µm. (G-I) Effects of shCOL and shCOL10A1+DDR2 on the migration ability of SW1990 and BXPC-3 cells by transwell assays. Scale bar, 50 µm. * P < 0.05, ** P < 0.01, **** P < 0.0001. ns, no significance.

Article Snippet: The following primary antibodies were used: DDR2, 1:200 (ABclonal); COL10A1, 1:200 (BOSTER); E-cadherin, N-cadherin, Vimentin, 1:200 (Cell Signaling Technology).

Techniques: Over Expression, Disruption, Tube Formation Assay, CCK-8 Assay, Migration

Journal: Frontiers in Oncology

Article Title: COL10A1-DDR2 axis promotes the progression of pancreatic cancer by regulating MEK/ERK signal transduction

doi: 10.3389/fonc.2022.1049345

Figure Lengend Snippet: COL10A1-DDR2 axis promotes EMT in PDAC cells through activation of MEK/ERK pathway. (A) GSEA validated EMT-related pathways in high COL10A1 expression PDAC cohorts of GSE15471 dataset. (B) The effect of COL10A1 on the expressions of EMT signaling proteins by Immunofluorescence. (C) The effect of COL10A1 and DDR2 on the expressions of EMT signaling proteins by Western blot. (D) Western blot detection of the effect of overexpression of DDR2 on protein expression changes in the EMT pathway induced by interference with COL10A1. (E) Western blot analyses of MEK/ERK pathway proteins in the indicated cells. (F) The protein expression of EMT pathway was detected by western blot in PDAC cells after treated with ERK inhibitor SCH772984. (G) H&E staining and IHC staining of COL10A1, DDR2, P-ERK, E-cadherin, N-cadherin and Vimentin in PDAC orthotopic models. Scale bars, 20 μm.

Article Snippet: The following primary antibodies were used: DDR2, 1:200 (ABclonal); COL10A1, 1:200 (BOSTER); E-cadherin, N-cadherin, Vimentin, 1:200 (Cell Signaling Technology).

Techniques: Activation Assay, Expressing, Immunofluorescence, Western Blot, Over Expression, Staining, Immunohistochemistry

Journal: Frontiers in Oncology

Article Title: COL10A1-DDR2 axis promotes the progression of pancreatic cancer by regulating MEK/ERK signal transduction

doi: 10.3389/fonc.2022.1049345

Figure Lengend Snippet: Model: COL10A1 activated the MEK/ERK signaling pathway of pancreatic cancer cells and promoted the EMT of pancreatic cancer cells through a DDR2-dependent pathway.

Article Snippet: The following primary antibodies were used: DDR2, 1:200 (ABclonal); COL10A1, 1:200 (BOSTER); E-cadherin, N-cadherin, Vimentin, 1:200 (Cell Signaling Technology).

Techniques:

Journal: medRxiv

Article Title: A brain-persistent DDR2-degrading antibody reverses Alzheimer’s pathologies by restoring brain fluid dynamics and metabolic clearance

doi: 10.64898/2026.03.17.26348575

Figure Lengend Snippet: (A) CBF assessed by LSCI in WT, APP/PS1, and astrocyte specific Ddr2 overexpressing APP/PS1 mice. Left: Representative pseudocolor perfusion maps. Right: Quantification of relative perfusion units (Mean ± SEM; ** p < 0.01, one-way ANOVA). (B) Correlation between CBF (from A) and cortical Ddr2 mRNA levels (qPCR) across individual mice (Pearson r = −0.6081, p < 0.01). (C) In vivo (top) and ex vivo (bottom) whole brain fluorescence imaging after intravenous injection of the DDR2 targeting probe 1A12-mCherry. (D) Validation of 1A12-mCherry brain delivery and target specificity: Ex vivo brain sections for intrinsic mCherry fluorescence (red), anti-His tag immunofluorescence staining (green), and anti-DDR2 antibody HL2 staining (purple), the areas outlined by white squares are magnified in the adjacent panels. Scale bar: 30 μm for original images and 10 μm for enlarged images. (E) Schematic of the sequential probe injection protocol for vascular perfusion assessment: 1A12-mCherry followed 40 min later by Dextran-FITC (70 kDa). (F) Whole brain fluorescence imaging of vascular perfusion with Dextran-FITC (70 kDa). (G) Two photon microscopy of cortical vasculature. Representative images show mCherry signal (red) and dextran-FITC vasculature (green) in WT, APP/PS1, and APP/PS1-DDR2 mice, scale bars: 50 μm. (H) Representative two-photon microscopy images of vascular leakage after injection of dextran-FITC (4 kDa, green) and dextran-RB (70 kDa, red), scale bars: 50 μm. (I) Ventricular morphology analyzed by MRI based volumetric reconstruction. Left: Representative images of lateral ventricles from each group. Right: Quantification of lateral ventricular volume (Mean ± SEM; * p < 0.05, ** p < 0.01, one-way ANOVA). (J) CSF flow assessed by cisterna magna injection of high molecular weight dextran (70 kDa, green). Fluorescence stereo microscope images displaying the spatial distribution of the glymphatic system (green) in the brains ( top row ), and corresponding whole brain fluorescence imaging system scans of the same brains( bottom row ), scale bars: 2 mm.

Article Snippet: The primary antibodies used in this study were as follows: anti-DDR2 (1:1000; Servicebio, GB112568), anti-DDR2 (1:1000, R&D, AF2538), anti-BACE1 (1:50; Cell Signaling Technology, no. 5606), anti-Collagen IV (1:1000; abcam, ab6586), anti-PDGFRβ (1:1000; Cell Signaling Technology, no. 3169), anti-Occludin (1:1000; Cell Signaling Technology, no. 91131), anti-ZO1 (1:1000; abcam, ab276131), anti-β-actin (1:1000; Yeasen, 30102ES60).

Techniques: In Vivo, Ex Vivo, Fluorescence, Imaging, Injection, Biomarker Discovery, Immunofluorescence, Staining, Microscopy, High Molecular Weight