Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: YAP Overexpression in Breast Cancer Cells Promotes Angiogenesis through Activating YAP Signaling in Vascular Endothelial Cells

doi: 10.1155/2022/5942379

Figure Lengend Snippet: Conditioned medium from YAP overexpressed breast cancer cells promoted tube formation, migration, and proliferation of human umbilical vein endothelial cells (HUVECs). (a) MDA-MB-231 cells were transfected with YAP CRISPR/Cas9 KO plasmid (YAP KD) and YAP CRISPR activation plasmids (YAP OE), YAP expression was detected by Western blot. (b) After starved from FBS for 16 hr, human umbilical vein endothelial cells (HUVECs) were treated with conditioned medium obtained from YAP knock down (CM-YAP-) and overexpressed (CM-YAP+) breast cancer cells for 12 hr. The total segment length and total branching length from tube formation assay are quantified with means ± SD from six independent representative fields. ∗∗∗ p < 0.001. (c) Migration of HUVECs with different conditioned medium treatment was conducted by Transwell assay. The results are from three independent experiments. ∗∗ p < 0.01, ∗∗∗ p < 0.001. (d) HUVECs were treated with different conditioned medium. Cell proliferation was tested by plate cloning formation. ∗∗ p < 0.01. The cloning formation rate is quantified with mean ± SD from six independent wells. ∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Cells were transfected with YAP CRISPR/Cas9 KO plasmid and YAP CRISPR activation plasmids using UltraCruz® Transfection Reagent (Santa Cruz, USA) according to manufacturer's instruction.

Techniques: Migration, Transfection, CRISPR, Plasmid Preparation, Activation Assay, Expressing, Western Blot, Knockdown, Tube Formation Assay, Transwell Assay, Cloning

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: YAP Overexpression in Breast Cancer Cells Promotes Angiogenesis through Activating YAP Signaling in Vascular Endothelial Cells

doi: 10.1155/2022/5942379

Figure Lengend Snippet: YAP expression in HUVECs involved in mediating CM-YAP+ induced tube formation, migration, and proliferation of HUVECs. (a) HUVECs were transfected with YAP CRISPR/Cas9 KO plasmid (YAP KD), YAP expression was detected by Western blot. (b) After starved from FBS for 16 hr, HUVECs, and YAP knock down (KD) HUVECs were treated with different conditioned medium obtained from breast cancer cells for 12 hr. The total segment length and total branching length are quantified with mean ± SD from six independent representative fields ∗ p < 0.05, ∗∗ p < 0.01. (c) Migration of HUVECs and YAP knock down HUVECs with different conditioned medium treatment was conducted by Transwell assay. The results are from three independent experiments, ∗∗ p < 0.01. (d) HUVECs and YAP KD HUVECs were treated with different conditioned medium. Cell proliferation was tested by plate cloning formation. The cloning formation rate is quantified with mean ± SD from six independent wells. ∗∗ p < 0.01.

Article Snippet: Cells were transfected with YAP CRISPR/Cas9 KO plasmid and YAP CRISPR activation plasmids using UltraCruz® Transfection Reagent (Santa Cruz, USA) according to manufacturer's instruction.

Techniques: Expressing, Migration, Transfection, CRISPR, Plasmid Preparation, Western Blot, Knockdown, Transwell Assay, Cloning

Journal: Translational Cancer Research

Article Title: IGF2BP3 -mediated stabilization of RHOA mRNA promotes osteosarcoma proliferation via Hippo signaling

doi: 10.21037/tcr-2025-1-2626

Figure Lengend Snippet: The inhibitory effect of OS cells triggered by IGF2BP3 knockdown was reversed by RHOA overexpression. (A) Protein levels of RHOA in 143B and U2OS cells transfected with IGF2BP3 knockdown, RHOA overexpression or negative control. (B,C) RHOA overexpression partly reversed the suppressed effects of IGF2BP3 knockdown on the cell viability of 143B and U2OS cells using CCK-8 assay. (D) The decrease in the proportion of EdU-positive OS cells due to IGF2BP3 knockdown was partly reversed by RHOA overexpression. (E,F) RHOA overexpression partly reversed the suppressed effects of IGF2BP3 knockdown on the colony-formation capability of 143B and U2OS cells (crystal violet staining; magnification, 1×). (G,H) RHOA overexpression partly reversed the induced effect of IGF2BP3 knockdown on the apoptosis of OS cells. Results are displayed as mean ± SD; *, P<0.05; **, P<0.01; ***, P<0.001. CCK-8, Cell Counting Kit-8; CON, control; EdU, 5-ethynyl-2'-deoxyuridine; FITC, fluorescein isothiocyanate; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; IGF2BP3, insulin-like growth factor 2 mRNA-binding protein 3; mRNA, messenger RNA; RHOA, Ras homolog family member A; OS, osteosarcoma; PI, propidium iodide; SD, standard deviation; sh, short hairpin RNA.

Article Snippet: Additionally, recombinant lentiviral vectors carrying RHOA and YAP overexpression constructs were purchased from OBiO Technology Corp. (Shanghai, China).

Techniques: Knockdown, Over Expression, Transfection, Negative Control, CCK-8 Assay, Staining, Cell Counting, Control, Binding Assay, Standard Deviation, shRNA

Journal: Translational Cancer Research

Article Title: IGF2BP3 -mediated stabilization of RHOA mRNA promotes osteosarcoma proliferation via Hippo signaling

doi: 10.21037/tcr-2025-1-2626

Figure Lengend Snippet: IGF2BP3 stimulates OS progression via RHOA/YAP pathway. (A) Protein levels of phosphorylation, as well as the total levels of YAP and LATS1 following the overexpression of RHOA in IGF2BP3 -silenced cells. (B) WB confirmation of YAPS127A (a constitutively active form of YAP that cannot be phosphorylated by LATS kinase) overexpression in sh- IGF2BP3 OS cells. (C,D) YAPS127A expression partly reversed the suppressed effects of IGF2BP3 knockdown on the cell viability of 143B and U2OS cells using CCK-8 assay. (E,F) The EdU-positive cell reduction effect caused by IGF2BP3 knockdown was partially reversed by YAPS127A overexpression. (G,H) The overexpression of YAPS127A partly reversed the suppressed effects of IGF2BP3 knockdown on the colony-formation capability of 143B and U2OS cells. (I,J) Overexpression of YAPS127A partly reversed the induced effect of IGF2BP3 knockdown on the apoptosis of OS cells. Results are displayed as mean ± SD; **, P<0.01; ***, P<0.001. CCK-8, Cell Counting Kit-8; CON, control; EdU, 5-ethynyl-2'-deoxyuridine; FITC, fluorescein isothiocyanate; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; IGF2BP3, insulin-like growth factor 2 mRNA-binding protein 3; mRNA, messenger RNA; RHOA, Ras homolog family member A; OS, osteosarcoma; p-, phosphorylated-; PI, propidium iodide; SD, standard deviation; sh, short hairpin RNA; YAP, yes-associated protein.

Article Snippet: Additionally, recombinant lentiviral vectors carrying RHOA and YAP overexpression constructs were purchased from OBiO Technology Corp. (Shanghai, China).

Techniques: Phospho-proteomics, Over Expression, Expressing, Knockdown, CCK-8 Assay, Cell Counting, Control, Binding Assay, Standard Deviation, shRNA

Journal: iScience

Article Title: CircPOLA2 sensitizes non-small cell lung cancer cells to ferroptosis and suppresses tumorigenesis via the Merlin-YAP signaling pathway

doi: 10.1016/j.isci.2024.110832

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti-YAP , Cell Signaling Technology , Cat#14074; RRID: AB_2650491.

Techniques: Control, Virus, Recombinant, Magnetic Beads, Protease Inhibitor, Silver Staining, Multiple Displacement Amplification, Purification, Cell Viability Assay, CCK-8 Assay, In Vitro, Sequencing, Expressing, Plasmid Preparation, Cloning, Software

Journal: Cancer research

Article Title: Recycling endosomes in mature epithelia restrain tumorigenic signaling

doi: 10.1158/0008-5472.CAN-18-4075

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit anti – YAP/TAZ (D24E4) , Cell Signaling , 8418, RRID:AB_10950494.

Techniques: Plasmid Preparation

Journal: Cell Reports

Article Title: YAP Activity Is Necessary and Sufficient for Basal Progenitor Abundance and Proliferation in the Developing Neocortex

doi: 10.1016/j.celrep.2019.03.091

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-phospho-YAP-S127 (1:50) , Cell Signaling , Cat# 4911; RRID: AB_2218913.

Techniques: Recombinant, Imaging, Plasmid Preparation, CRISPR, Software