Journal: International Journal of Oncology

Article Title: Fibulin-4 promotes osteosarcoma invasion and metastasis by inducing epithelial to mesenchymal transition via the PI3K/Akt/mTOR pathway

doi: 10.3892/ijo.2017.3921

Figure Lengend Snippet: Expression levels of fibulin-4, CYR61 and ALP in human osteosarcoma tissues. (A) Fibulin-4 expression levels in osteosarcoma tissue, fibrous dysplasia tissue and normal tissue were measured by IHC. (B) ALP expression levels in osteosarcoma tissue, fibrous dysplasia tissue and normal tissue were measured by IHC. (C) CYR61 expression levels in osteosarcoma tissue, fibrous dysplasia tissue and normal tissue were measured by IHC. (D) According to the Pearson's product-moment correlation coefficient, the expression of fibulin-4 positively correlated with CYR61 and ALP, using photodensities detected by software Image-Pro Plus 6.0. (Magnification, ×200).

Article Snippet: All sections and coverslips were treated with 3% hydrogen peroxide (H 2 O 2 ) and goat serum for 30 min sequentially, to block endogenous peroxidase and the non-specific binding sites, and then incubated with rabbit anti-human fibulin-4, alkaline phosphatase (ALP), and cysteine-rich angiogenic inducer 61 (CYR61) antibodies (ab125073, ab108337, ab24448; Abcam) at working dilutions of 1:200 overnight at 4°C.

Techniques: Expressing, Software

Journal: International Journal of Oncology

Article Title: Fibulin-4 promotes osteosarcoma invasion and metastasis by inducing epithelial to mesenchymal transition via the PI3K/Akt/mTOR pathway

doi: 10.3892/ijo.2017.3921

Figure Lengend Snippet: Effects of fibulin-4 knockdown and overexpression on EMT genes, CYR61 and ALP, which were correlated to tumor progression. After lentivirus transfections, EMT markers, including E-cadherin, N-cadherin, vimentin, Snail, Slug and Twist were measured by (A) western blotting and (B) real-time q-RT-PCR in the lentivirus transfection systems of the differently invasive osteosarcoma cell subclones. (C) EMT markers (E-cadherin, N-cadherin, vimentin, Snail, Slug and Twist), CYR61 and ALP were detected by western blotting in fibulin-4 shRNA-infected HOS cells and pLVX-fibulin-4-infected U-2OS cells. Fibulin-4 knockdown significantly increased the expression of E-cadherin, and decreased the expression of N-cadherin, vimentin, Snail, Slug and Twist; whereas fibulin-4 upregulation decreased the expression of E-cadherin, and increased the expression of N-cadherin, vimentin, Snail, Slug and Twist. CYR61 and ALP were downregulated following fibulin-4 shRNA infection, in association with the knockdown of fibulin-4 expression, moreover, with fibulin-4 upregulation, CYR61 and ALP were increased following pLVX-fibulin-4 infection. * P<0.05.

Article Snippet: All sections and coverslips were treated with 3% hydrogen peroxide (H 2 O 2 ) and goat serum for 30 min sequentially, to block endogenous peroxidase and the non-specific binding sites, and then incubated with rabbit anti-human fibulin-4, alkaline phosphatase (ALP), and cysteine-rich angiogenic inducer 61 (CYR61) antibodies (ab125073, ab108337, ab24448; Abcam) at working dilutions of 1:200 overnight at 4°C.

Techniques: Over Expression, Transfection, Western Blot, Reverse Transcription Polymerase Chain Reaction, shRNA, Infection, Expressing

Journal: Investigative Ophthalmology & Visual Science

Article Title: Downregulation of microRNA-224-3p Hampers Retinoblastoma Progression via Activation of the Hippo-YAP Signaling Pathway by Increasing LATS2

doi: 10.1167/iovs.61.3.32

Figure Lengend Snippet: Primer Sequences for RT-qPCR

Article Snippet: The primary antibodies, including LATS2 (1 µg/mL, ab110780), B-cell lymphoma 2 (Bcl-2) (1:1000, ab32124), Bcl-2 associated X protein (Bax) (1:1000, ab3250), vascular endothelial growth factor (VEGF) (1:1000, ab32152), tafazzin (TAZ) (1 µg/mL, ab84927), Yes associated protein (YAP) (1:5000, ab52771), p-YAP (1:10000, ab76252), connective tissue growth factor (CTGF) (1:1000, ab6992), and cysteine rich angiogenic inducer 61 (CYR61) (1 µg/mL, ab24448), were purchased from Abcam, Inc. (Cambridge, UK) except p-TAZ (1:1000, sc-17610-R; Santa Cruz, USA).

Techniques: Sequencing

Journal: Investigative Ophthalmology & Visual Science

Article Title: Downregulation of microRNA-224-3p Hampers Retinoblastoma Progression via Activation of the Hippo-YAP Signaling Pathway by Increasing LATS2

doi: 10.1167/iovs.61.3.32

Figure Lengend Snippet: miR-224-3p inhibits the Hippo-YAP signaling pathway by downregulating expression of LATS2. ( A ) Involvement of LATS2 in the Hippo-YAP signaling pathway analyzed on KEGG. ( B ) mRNA expression of TAZ and YAP in cells determined by PT-qPCR. ( C ) Protein expression of TAZ, YAP, p-TAZ, and p-YAP in cells normalized to GAPDH determined by Western blot analysis. ( D ) mRNA expression of TAZ, YAP, CTGF, and CYR61 in cells determined by PT-qPCR. ( E ) Protein expression of TAZ, YAP, CTGF, CYR61, p-TAZ, and p-YAP in cells normalized to GAPDH determined by Western blot analysis. * P < 0.05 versus Y79 cells transfected with NC mimic or oe-NC; # P < 0.05 versus Y79 cells transfected with NC inhibitor or cotransfected with miR-224-3p mimic and oe-NC. Data expressed by mean ± standard deviation among multiple groups were analyzed using 1-way ANOVA, followed by Tukey's post hoc test. The experiment was repeated three times independently.

Article Snippet: The primary antibodies, including LATS2 (1 µg/mL, ab110780), B-cell lymphoma 2 (Bcl-2) (1:1000, ab32124), Bcl-2 associated X protein (Bax) (1:1000, ab3250), vascular endothelial growth factor (VEGF) (1:1000, ab32152), tafazzin (TAZ) (1 µg/mL, ab84927), Yes associated protein (YAP) (1:5000, ab52771), p-YAP (1:10000, ab76252), connective tissue growth factor (CTGF) (1:1000, ab6992), and cysteine rich angiogenic inducer 61 (CYR61) (1 µg/mL, ab24448), were purchased from Abcam, Inc. (Cambridge, UK) except p-TAZ (1:1000, sc-17610-R; Santa Cruz, USA).

Techniques: Expressing, Western Blot, Transfection, Standard Deviation

Journal: Investigative Ophthalmology & Visual Science

Article Title: Downregulation of microRNA-224-3p Hampers Retinoblastoma Progression via Activation of the Hippo-YAP Signaling Pathway by Increasing LATS2

doi: 10.1167/iovs.61.3.32

Figure Lengend Snippet: A mechanism map depicting the role of the miR-224-3p/LATS2/Hippo-YAP axis in the progression of retinoblastoma. miR-224-3p targets and negatively regulates LATS2 to inhibit the YAP/TAZ phosphorylation, whereby the Hippo signaling pathway activation is inhibited; following that, the downstream genes CTGF and CYR61 are upregulated, by which the apoptosis of retinoblastoma cells is suppressed while proliferation and angiogenesis are promoted.

Article Snippet: The primary antibodies, including LATS2 (1 µg/mL, ab110780), B-cell lymphoma 2 (Bcl-2) (1:1000, ab32124), Bcl-2 associated X protein (Bax) (1:1000, ab3250), vascular endothelial growth factor (VEGF) (1:1000, ab32152), tafazzin (TAZ) (1 µg/mL, ab84927), Yes associated protein (YAP) (1:5000, ab52771), p-YAP (1:10000, ab76252), connective tissue growth factor (CTGF) (1:1000, ab6992), and cysteine rich angiogenic inducer 61 (CYR61) (1 µg/mL, ab24448), were purchased from Abcam, Inc. (Cambridge, UK) except p-TAZ (1:1000, sc-17610-R; Santa Cruz, USA).

Techniques: Activation Assay