Journal: Stem Cells International

Article Title: Endothelial Cells Promote Migration of Mesenchymal Stem Cells via PDGF-BB/PDGFR β -Src-Akt in the Context of Inflammatory Microenvironment upon Bone Defect

doi: 10.1155/2022/2401693

Figure Lengend Snippet: Reagent information.

Article Snippet: JNJ-10198409 , PDGFR β , 5 μ M , 1 h , Pre-treat MSCs , MedChemExpress, Monmouth Junction, NJ, USA.

Techniques: Concentration Assay, Injection

Journal: Stem Cells International

Article Title: Endothelial Cells Promote Migration of Mesenchymal Stem Cells via PDGF-BB/PDGFR β -Src-Akt in the Context of Inflammatory Microenvironment upon Bone Defect

doi: 10.1155/2022/2401693

Figure Lengend Snippet: MSCs migrated toward ECs via PDGF-BB/PDGFR β in the inflammatory microenvironment. (a) Representative images of migrated hBMSCs that had received different pre-treatments or had been exposed to different inducing media. The migration capacity of hBMSCs was determined using a Transwell culture system. The quantification of migrated cells was shown as a bar graph. Scale bar, 50 μ m. ∗ P < 0.05. (b) Representative images of wound healing assays. The rate of scratch wound closure was shown as a bar graph. Scale bar, 200 μ m. ∗ P < 0.05. EGM: endothelial cell growth medium-2. EC-CM: conditioned media of endothelial cells. IEC-CM: conditioned media of endothelial cells in the context of inflammatory microenvironment; shPDGF-BB: short hairpin RNA targeting pdgfb ; shPDGFR β : shRNA targeting pdgfrb .

Article Snippet: JNJ-10198409 , PDGFR β , 5 μ M , 1 h , Pre-treat MSCs , MedChemExpress, Monmouth Junction, NJ, USA.

Techniques: Migration, shRNA

Journal: Stem Cells International

Article Title: Endothelial Cells Promote Migration of Mesenchymal Stem Cells via PDGF-BB/PDGFR β -Src-Akt in the Context of Inflammatory Microenvironment upon Bone Defect

doi: 10.1155/2022/2401693

Figure Lengend Snippet: Src and Akt functioned downstream of PDGFR β . (a) Representative images of migrated hBMSCs in Transwell culture systems. The quantification of migrated cells was shown as a bar graph. Data were compared with the group of IEC-CM from . Scale bar, 50 μ m. ∗ P < 0.05. (b) Representative images of wound healing assays. The rate of scratch wound closure was shown as a bar graph. Data were compared with the group of IEC-CM from . Scale bar, 200 μ m. ∗ P < 0.05. IEC-CM: conditioned media of endothelial cells in the context of inflammatory microenvironment; shPDGFR β : short hairpin RNA targeting pdgfrb ; shSrc: shRNA targeting src ; shAkt: shRNA targeting akt .

Article Snippet: JNJ-10198409 , PDGFR β , 5 μ M , 1 h , Pre-treat MSCs , MedChemExpress, Monmouth Junction, NJ, USA.

Techniques: shRNA

Journal: Stem Cells International

Article Title: Endothelial Cells Promote Migration of Mesenchymal Stem Cells via PDGF-BB/PDGFR β -Src-Akt in the Context of Inflammatory Microenvironment upon Bone Defect

doi: 10.1155/2022/2401693

Figure Lengend Snippet: Src bridged connection between PDGFR β and Akt during ECs-induced MSCs migration. (a) Gene and protein expression of Src in migrating hBMSCs. (b) Gene and protein expression of Akt in migrating hBMSCs. EC-CM: conditioned media of ECs; IEC-CM: conditioned media of ECs in the context of inflammatory microenvironment; shPDGFR β : short hairpin RNA targeting pdgfrb ; shSrc: shRNA targeting src ; shAkt: shRNA targeting akt . ∗ P < 0.05.

Article Snippet: JNJ-10198409 , PDGFR β , 5 μ M , 1 h , Pre-treat MSCs , MedChemExpress, Monmouth Junction, NJ, USA.

Techniques: Migration, Expressing, shRNA

Journal: Cancer Research

Article Title: Discovery and Evaluation of Biomarkers for Triple-Negative Breast Cancer Subtypes Uncovers Patient Stratification and Targeted Therapeutic Strategies

doi: 10.1158/0008-5472.CAN-24-2758

Figure Lengend Snippet: TAGL as a predictive marker for dasatinib sensitivity. A, Western blot of control and TAGLN -KO cells in Hs578T (top) and BT-549 (bottom) cells. B, Representative immunofluorescence of Hs578T and BT549 control and TAGLN -KO cells, stained with αTAGL (green) and DAPI (blue). Scale bar, 50 μm. C, Cell proliferation in control (black) and TAGLN -KO (purple) cells measured by MTT assays over 72 hours. D, Relative percentage of migrated cells in control and TAGLN -KO cells, assessed by transwell assays. In C and D , values represent the mean ± SEM of at least three experimental replicates, and statistical significance was determined using one-way ANOVA. E, Drug–response curves for cell viability of control and TAGLN -KO cells treated with dasatinib at increasing concentrations. F, Western blot of known dasatinib targets in control and TAGLN -KO cells. G, Data from the DepMap portal showing the correlation between TAGLN and PDGFRB in breast cancer cell lines. H, Western blot of PDGFRβ in TAGLN -KO and cells and clones constitutively expressing PDGFRB . I, Drug–response curves for cell viability of TAGLN -KO and TAGLN -KO/ PBGFRB cells treated with dasatinib at increasing concentrations. In E and I , solid lines represent the mean of three biological replicates performed in technical replicates. The dashed line indicates their IC 50 value. **, P < 0.01; ****, P < 0.0001; ns, not significant.

Article Snippet: For PDGFRB overexpression, lentiviral particles were generated using the PDGFRB Human Tagged Lenti ORF Clone (RC206377L4, OriGene) plasmid and used to transduce target cells.

Techniques: Marker, Western Blot, Control, Immunofluorescence, Staining, Clone Assay, Expressing

Journal: Journal of Cancer

Article Title: P68 RNA Helicase facilitates Breast Cancer progression by promoting Proliferation and Migration via PDGFR-β/AR axis.

doi: 10.7150/jca.61505

Figure Lengend Snippet: Figure 3. P68 RNA helicase transcriptionally regulates PDGFR-β expression in breast cancer cells. (A, B) Levels of p-PDGFRβ (IB: p-PDGFRβ) and pERK ½ (IB:pERK1/2), an established effector of PDGFR-β signaling in MDAMB231 (A) and BT549 (B) cells were analyzed by immunoblot upon PDGF-BB (20 ng/ml, 24h) stimulation. Levels of ERK ½ (IB:ERK1/2) and PDGFR-β (IB: PDGFRβ) are the loading controls. (C, D) Immunoblot analysis (C) of PDGFR-β (IB: PDGFR−β), EGFR (IB:EGFR), and FGFR (IB:FGFR) and their relative mRNA expression (D) in PDGF-BB stimulated MDAMB231 and BT549 cells upon p68 knockdown. Knockdown efficiency of p68 (IB:p68) was analyzed by immunoblot. Immunoblot of β-actin (IB:β-actin) is a loading control. (E) In vitro scratch wound healing assay of MDAMB231 cells transfected with p68 siRNA or co-transfected with p68 siRNA and PDGFRβ plasmid compared with control siRNA at 0 h and 24 h upon PDGF-BB stimulation. (F & G) Quantitative analyses of the migrating PDGF-BB stimulated MDAMB231 (F), and BT549 (G). The results represent findings of five independent experiments. Error bars represent mean ± S.E.M. **P<0.01, ****P<0.0001, ns denotes non-significant.

Article Snippet: To overexpress PDGFR−β in MDAMB231 and BT549 cells, 2.5 μg of PDGFR-β plasmid (Origene RC206377) was transfected for 48 h using LipofectamineTM 3000 Transfection Reagent kit (Thermo Fisher scientific).

Techniques: Expressing, Western Blot, Knockdown, Control, In Vitro, Wound Healing Assay, Transfection, Plasmid Preparation

Journal: Journal of Cancer

Article Title: P68 RNA Helicase facilitates Breast Cancer progression by promoting Proliferation and Migration via PDGFR-β/AR axis.

doi: 10.7150/jca.61505

Figure Lengend Snippet: Figure 5. PDGFR-β regulates AR expression and mediates the effect of DHT in proliferation of breast cancer cells. (A & B) Immunoblot analysis (A), and relative mRNA expression (B) of AR in PDGF-BB stimulated MDAMB231 and BT549 cells upon PDGFR-β knockdown. Knockdown efficiency of PDGFR-β (IB: PDGFR-β) was analyzed by immunoblot. β-actin (IB: β-actin) is the loading control. (C) Cell proliferation of MDAMB231 and BT549 cells untreated (open bar) or treated (filled bar) with DHT (1 nM for 24 h) upon PDGFR-β knock down compared with the control. The experiments were performed in triplicate. Error bars represent mean ± S.E.M. **P<0.01, ***P<0.001, ns denotes non-significant.

Article Snippet: To overexpress PDGFR−β in MDAMB231 and BT549 cells, 2.5 μg of PDGFR-β plasmid (Origene RC206377) was transfected for 48 h using LipofectamineTM 3000 Transfection Reagent kit (Thermo Fisher scientific).

Techniques: Expressing, Western Blot, Knockdown, Control

Journal: Journal of Cancer

Article Title: P68 RNA Helicase facilitates Breast Cancer progression by promoting Proliferation and Migration via PDGFR-β/AR axis.

doi: 10.7150/jca.61505

Figure Lengend Snippet: Figure 6. Nuclear PDGFR-β regulates AR expression in breast cancer cells. (A) Representative images of immunofluorescence staining of PDGFR-β (red) MDAMB231 cells treated with PDGFR-β inhibitor, AG1296 with or without PDGF-BB stimulation for 24 h compared with the control. Nuclei were stained with Hoescht (blue). (B) Levels of AR (IB: AR) in MDMB231 cells upon indicated conditions were analyzed by immunoblot. Levels of pPDGFR-β (IB: pPDGFR-β), PDGFR-β (IB:PDGFR-β) and p68 (IB:p68) are the controls, indicating the efficiency of AG1296, PDGFR-β knockdown efficiency, and total p68 respectively. Immunoblot of β-actin (IB:β-actin) is the loading control. (C) Schematic illustration of p68- PDGFRβ-AR axis in breast cancer cells upon PDGF-BB stimulation.

Article Snippet: To overexpress PDGFR−β in MDAMB231 and BT549 cells, 2.5 μg of PDGFR-β plasmid (Origene RC206377) was transfected for 48 h using LipofectamineTM 3000 Transfection Reagent kit (Thermo Fisher scientific).

Techniques: Expressing, Immunofluorescence, Staining, Control, Western Blot, Knockdown