anti cysteine rich angiogenic inducer 61  (Abcam)

Journal: Cellular and Molecular Life Sciences

Article Title: YAP1-CPNE3 positive feedback pathway promotes gastric cancer cell progression

doi: 10.1007/s00018-024-05178-3

Figure Lengend Snippet: Screening of CPNE3 as a downstream target gene regulated by YAP1. A mRNA sequencing in BGC-823 cells with downregulated YAP1 expression. B Examination of mRNA expression by quantitative reverse transcription polymerase chain reaction (RT-qPCR) after downregulating YAP1 in BGC-823 cells. C , D mRNA expression levels of YAP1 , CPNE3 , and CYR61 in AGS and HGC-27 cells (transfected with NC, siYAP1-#1, and siYAP1-#2 siRNAs). E Two-dimensional visualization of CPNE3 and YAP1 in single-cell clusters in patients with gastric cancer (GC). F Protein expression levels of YAP1, CPNE3, and CYR61 following down-regulation of YAP1 expression in AGS and HGC-27 cells. G , H After gradient overexpression of Flag-YAP1, the YAP1, CPNE3, and CYR61 proteins and mRNA levels were detected by western blotting (WB) and RT-qPCR, respectively. Three independent biological experiments were performed, and statistical significance is denoted by * p < 0.05 and ** p < 0.01

Article Snippet: IHC was performed with a normal inverted microscope (NION, Japan) by using an anti-YAP1 antibody (1:200; Cell Signaling Technology), anti-CPNE3 antibody (1:50; abcam, Proteintech), anti-cysteine-rich angiogenic inducer 61 (CYR61) antibody (1:100; Proteintech), and anti-RAD51 antibody (1:100; Proteintech).

Techniques: Sequencing, Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Transfection, Over Expression, Western Blot

Journal: Cellular and Molecular Life Sciences

Article Title: YAP1-CPNE3 positive feedback pathway promotes gastric cancer cell progression

doi: 10.1007/s00018-024-05178-3

Figure Lengend Snippet: CPNE3 is associated with the Hippo-YAP1 pathway in GC. A Enrichment analysis of CPNE3 with Hippo-YAP1. B Pearson correlation analyses of CPNE3 with Hippo-YAP1 C – E Using siRNA, the expression of CPNE3 at the mRNA and protein levels were downregulated in BGC-823 and MKN-28 cells. F , G Up-regulation of CPNE3 expression by transfection of HA-CPNE3 plasmid in AGS and HGC-27 cells. H WB revealed that the expression of YAP1 and its target gene CYR61 were both considerably upregulated in AGS and HGC-27 cells using the same technique used to boost CPNE3 expression as above. I , J Confocal images of HA-CPNE3 plasmid transfection in HGC-27 cells and siCPNE3-#1 or siCPNE3-#2 transfection in BGC-823 cells with YAP1 labeling

Article Snippet: IHC was performed with a normal inverted microscope (NION, Japan) by using an anti-YAP1 antibody (1:200; Cell Signaling Technology), anti-CPNE3 antibody (1:50; abcam, Proteintech), anti-cysteine-rich angiogenic inducer 61 (CYR61) antibody (1:100; Proteintech), and anti-RAD51 antibody (1:100; Proteintech).

Techniques: Expressing, Transfection, Plasmid Preparation, Labeling

Journal: Cellular and Molecular Life Sciences

Article Title: YAP1-CPNE3 positive feedback pathway promotes gastric cancer cell progression

doi: 10.1007/s00018-024-05178-3

Figure Lengend Snippet: CPNE3 promotes GC progression in a partial YAP1-dependent manner. BGC-823 and MKN-28 cells were treated with the ShCPNE3-#2 plasmid to downregulate the expression of CPNE3 and the Flag-YAP1 plasmid to concurrently increase the expression of YAP1. A WB was used to measure the expression of CPNE3, YAP1, and CYR61. B – G Phenotyping assays were used to determine the degree to which the CPNE3 depletion-induced suppression of GC cell proliferation, migration, invasion, colony formation, and drug resistance could be reversed by the overexpression of exogenous YAP1. H – K The capacity of MKN-45 cells to proliferate, invade, migrate, and form colonies was weakly inhibited by downregulation of CPNE3 expression in MKN-45 cells lacking YAP1 expression. ShYAP1-#1 plasmid-mediated stable YAP1 knockdown or HA-CPNE3 plasmid-mediated simultaneous overexpression of CPNE3 in AGS and HGC-27 cells. L WB was used to investigate the relevant protein levels. M – P The overexpression of CPNE3 did not alleviate the suppression of GC cell proliferation, migration, invasion, and colony formation brought on by the downregulation of YAP1. Three independent biological experiments were conducted, and statistical significance is shown by the notations, * p < 0.05 and ** p < 0.01

Article Snippet: IHC was performed with a normal inverted microscope (NION, Japan) by using an anti-YAP1 antibody (1:200; Cell Signaling Technology), anti-CPNE3 antibody (1:50; abcam, Proteintech), anti-cysteine-rich angiogenic inducer 61 (CYR61) antibody (1:100; Proteintech), and anti-RAD51 antibody (1:100; Proteintech).

Techniques: Plasmid Preparation, Expressing, Migration, Over Expression

Journal: Cellular and Molecular Life Sciences

Article Title: YAP1-CPNE3 positive feedback pathway promotes gastric cancer cell progression

doi: 10.1007/s00018-024-05178-3

Figure Lengend Snippet: CPNE3 promotes GC growth in vivo. A , B Cell line with stable silencing of CPNE3 expression was established in BGC-823 cells by infection with lentivirus encoding sgRNA targeting CPNE3 or negative control. C – E Silencing of CPNE3 significantly reduced tumor growth in vivo, and the weight and volume of the tumor tissues were significantly lower than those in controls (mean ± standard error of the mean [SEM] n = 10/group). F , G Immunohistochemistry (IHC) and WB tests were used to assess the protein levels of CPNE3, YAP1, and CYR61 in tumor tissues from subcutaneous cell line-derived xenograft models constructed from BGC-823 cells with CPNE3 expression downregulated by lentivirus. H – J Using the same method described above to validate the function of CPNE3 in a patient-derived xenograft (PDX) model of GC, silencing CPNE3 significantly reduced tumor growth, weight, and volume in vivo (mean ± SEM n = 6/group). K , L Protein expression of CPNE3, YAP1, and CYR61 were detected using WB and IHC, assays in BGC-823 cells after stably downregulating CPNE3 expression in the PDX model

Article Snippet: IHC was performed with a normal inverted microscope (NION, Japan) by using an anti-YAP1 antibody (1:200; Cell Signaling Technology), anti-CPNE3 antibody (1:50; abcam, Proteintech), anti-cysteine-rich angiogenic inducer 61 (CYR61) antibody (1:100; Proteintech), and anti-RAD51 antibody (1:100; Proteintech).

Techniques: In Vivo, Expressing, Infection, Negative Control, Immunohistochemistry, Derivative Assay, Construct, Stable Transfection

Journal: Cellular and Molecular Life Sciences

Article Title: YAP1-CPNE3 positive feedback pathway promotes gastric cancer cell progression

doi: 10.1007/s00018-024-05178-3

Figure Lengend Snippet: CPNE3 is an independent prognostic factor that causes poor prognosis in patients with GC. A Using Western blotting, the protein levels of CPNE3 in eight pairs of GC patient tissue samples were measured. B CPNE3 protein expression was examined the IHC of tumor tissues (n = 20) and matched normal tissues (n = 20) from patients with GC. C – E Overall survival and CPNE3 expression were used to stratify the patients in the training, validation, and training + validation groups, followed by Kaplan–Meier analysis. F The chi-square test was used to determine the relationship between the expression of CPNE3 and that of YAP1, CYR61, and RAD51. G Kaplan–Meier survival analysis was performed in 100 patients who were categorized based on their CPNE3 and YAP1 protein levels. H Diagram by Figdraw showing the process through which the CPNE3-YAP1 positive feedback loop promotes GC

Article Snippet: IHC was performed with a normal inverted microscope (NION, Japan) by using an anti-YAP1 antibody (1:200; Cell Signaling Technology), anti-CPNE3 antibody (1:50; abcam, Proteintech), anti-cysteine-rich angiogenic inducer 61 (CYR61) antibody (1:100; Proteintech), and anti-RAD51 antibody (1:100; Proteintech).

Techniques: Western Blot, Expressing

Journal: Cellular and Molecular Life Sciences

Article Title: YAP1-CPNE3 positive feedback pathway promotes gastric cancer cell progression

doi: 10.1007/s00018-024-05178-3

Figure Lengend Snippet: Screening of CPNE3 as a downstream target gene regulated by YAP1. A mRNA sequencing in BGC-823 cells with downregulated YAP1 expression. B Examination of mRNA expression by quantitative reverse transcription polymerase chain reaction (RT-qPCR) after downregulating YAP1 in BGC-823 cells. C , D mRNA expression levels of YAP1 , CPNE3 , and CYR61 in AGS and HGC-27 cells (transfected with NC, siYAP1-#1, and siYAP1-#2 siRNAs). E Two-dimensional visualization of CPNE3 and YAP1 in single-cell clusters in patients with gastric cancer (GC). F Protein expression levels of YAP1, CPNE3, and CYR61 following down-regulation of YAP1 expression in AGS and HGC-27 cells. G , H After gradient overexpression of Flag-YAP1, the YAP1, CPNE3, and CYR61 proteins and mRNA levels were detected by western blotting (WB) and RT-qPCR, respectively. Three independent biological experiments were performed, and statistical significance is denoted by * p < 0.05 and ** p < 0.01

Article Snippet: IHC was performed with a normal inverted microscope (NION, Japan) by using an anti-YAP1 antibody (1:200; Cell Signaling Technology), anti-CPNE3 antibody (1:50; abcam, Proteintech), anti-cysteine-rich angiogenic inducer 61 (CYR61) antibody (1:100; Proteintech), and anti-RAD51 antibody (1:100; Proteintech).

Techniques: Sequencing, Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Transfection, Over Expression, Western Blot

Journal: Cellular and Molecular Life Sciences

Article Title: YAP1-CPNE3 positive feedback pathway promotes gastric cancer cell progression

doi: 10.1007/s00018-024-05178-3

Figure Lengend Snippet: CPNE3 is associated with the Hippo-YAP1 pathway in GC. A Enrichment analysis of CPNE3 with Hippo-YAP1. B Pearson correlation analyses of CPNE3 with Hippo-YAP1 C – E Using siRNA, the expression of CPNE3 at the mRNA and protein levels were downregulated in BGC-823 and MKN-28 cells. F , G Up-regulation of CPNE3 expression by transfection of HA-CPNE3 plasmid in AGS and HGC-27 cells. H WB revealed that the expression of YAP1 and its target gene CYR61 were both considerably upregulated in AGS and HGC-27 cells using the same technique used to boost CPNE3 expression as above. I , J Confocal images of HA-CPNE3 plasmid transfection in HGC-27 cells and siCPNE3-#1 or siCPNE3-#2 transfection in BGC-823 cells with YAP1 labeling

Article Snippet: IHC was performed with a normal inverted microscope (NION, Japan) by using an anti-YAP1 antibody (1:200; Cell Signaling Technology), anti-CPNE3 antibody (1:50; abcam, Proteintech), anti-cysteine-rich angiogenic inducer 61 (CYR61) antibody (1:100; Proteintech), and anti-RAD51 antibody (1:100; Proteintech).

Techniques: Expressing, Transfection, Plasmid Preparation, Labeling

Journal: Cellular and Molecular Life Sciences

Article Title: YAP1-CPNE3 positive feedback pathway promotes gastric cancer cell progression

doi: 10.1007/s00018-024-05178-3

Figure Lengend Snippet: CPNE3 promotes GC progression in a partial YAP1-dependent manner. BGC-823 and MKN-28 cells were treated with the ShCPNE3-#2 plasmid to downregulate the expression of CPNE3 and the Flag-YAP1 plasmid to concurrently increase the expression of YAP1. A WB was used to measure the expression of CPNE3, YAP1, and CYR61. B – G Phenotyping assays were used to determine the degree to which the CPNE3 depletion-induced suppression of GC cell proliferation, migration, invasion, colony formation, and drug resistance could be reversed by the overexpression of exogenous YAP1. H – K The capacity of MKN-45 cells to proliferate, invade, migrate, and form colonies was weakly inhibited by downregulation of CPNE3 expression in MKN-45 cells lacking YAP1 expression. ShYAP1-#1 plasmid-mediated stable YAP1 knockdown or HA-CPNE3 plasmid-mediated simultaneous overexpression of CPNE3 in AGS and HGC-27 cells. L WB was used to investigate the relevant protein levels. M – P The overexpression of CPNE3 did not alleviate the suppression of GC cell proliferation, migration, invasion, and colony formation brought on by the downregulation of YAP1. Three independent biological experiments were conducted, and statistical significance is shown by the notations, * p < 0.05 and ** p < 0.01

Article Snippet: IHC was performed with a normal inverted microscope (NION, Japan) by using an anti-YAP1 antibody (1:200; Cell Signaling Technology), anti-CPNE3 antibody (1:50; abcam, Proteintech), anti-cysteine-rich angiogenic inducer 61 (CYR61) antibody (1:100; Proteintech), and anti-RAD51 antibody (1:100; Proteintech).

Techniques: Plasmid Preparation, Expressing, Migration, Over Expression

Journal: Cellular and Molecular Life Sciences

Article Title: YAP1-CPNE3 positive feedback pathway promotes gastric cancer cell progression

doi: 10.1007/s00018-024-05178-3

Figure Lengend Snippet: CPNE3 promotes GC growth in vivo. A , B Cell line with stable silencing of CPNE3 expression was established in BGC-823 cells by infection with lentivirus encoding sgRNA targeting CPNE3 or negative control. C – E Silencing of CPNE3 significantly reduced tumor growth in vivo, and the weight and volume of the tumor tissues were significantly lower than those in controls (mean ± standard error of the mean [SEM] n = 10/group). F , G Immunohistochemistry (IHC) and WB tests were used to assess the protein levels of CPNE3, YAP1, and CYR61 in tumor tissues from subcutaneous cell line-derived xenograft models constructed from BGC-823 cells with CPNE3 expression downregulated by lentivirus. H – J Using the same method described above to validate the function of CPNE3 in a patient-derived xenograft (PDX) model of GC, silencing CPNE3 significantly reduced tumor growth, weight, and volume in vivo (mean ± SEM n = 6/group). K , L Protein expression of CPNE3, YAP1, and CYR61 were detected using WB and IHC, assays in BGC-823 cells after stably downregulating CPNE3 expression in the PDX model

Article Snippet: IHC was performed with a normal inverted microscope (NION, Japan) by using an anti-YAP1 antibody (1:200; Cell Signaling Technology), anti-CPNE3 antibody (1:50; abcam, Proteintech), anti-cysteine-rich angiogenic inducer 61 (CYR61) antibody (1:100; Proteintech), and anti-RAD51 antibody (1:100; Proteintech).

Techniques: In Vivo, Expressing, Infection, Negative Control, Immunohistochemistry, Derivative Assay, Construct, Stable Transfection

Journal: Cellular and Molecular Life Sciences

Article Title: YAP1-CPNE3 positive feedback pathway promotes gastric cancer cell progression

doi: 10.1007/s00018-024-05178-3

Figure Lengend Snippet: CPNE3 is an independent prognostic factor that causes poor prognosis in patients with GC. A Using Western blotting, the protein levels of CPNE3 in eight pairs of GC patient tissue samples were measured. B CPNE3 protein expression was examined the IHC of tumor tissues (n = 20) and matched normal tissues (n = 20) from patients with GC. C – E Overall survival and CPNE3 expression were used to stratify the patients in the training, validation, and training + validation groups, followed by Kaplan–Meier analysis. F The chi-square test was used to determine the relationship between the expression of CPNE3 and that of YAP1, CYR61, and RAD51. G Kaplan–Meier survival analysis was performed in 100 patients who were categorized based on their CPNE3 and YAP1 protein levels. H Diagram by Figdraw showing the process through which the CPNE3-YAP1 positive feedback loop promotes GC

Article Snippet: IHC was performed with a normal inverted microscope (NION, Japan) by using an anti-YAP1 antibody (1:200; Cell Signaling Technology), anti-CPNE3 antibody (1:50; abcam, Proteintech), anti-cysteine-rich angiogenic inducer 61 (CYR61) antibody (1:100; Proteintech), and anti-RAD51 antibody (1:100; Proteintech).

Techniques: Western Blot, Expressing

Journal: Molecular Medicine Reports

Article Title: HOXB13 suppresses proliferation, migration and invasion, and promotes apoptosis of gastric cancer cells through transcriptional activation of VGLL4 to inhibit the involvement of TEAD4 in the Hippo signaling pathway

doi: 10.3892/mmr.2021.12361

Figure Lengend Snippet: HOXB13 inhibits the involvement of TEAD4 in Hippo signaling pathway by regulating VGLL4 expression. (A) Relative mRNA expression levels of VGLL4 in transfected HGC-27 cells were examined by RT-qPCR. ### P<0.001 vs. si-NC. (B) Protein and (C) mRNA expression levels of TEAD4 in HGC-27 cells were determined by western blot assay and RT-qPCR, respectively. **P<0.01, ***P<0.001 vs. Control; # P<0.05, ### P<0.001 vs. Ov-HOXB13 + si-NC. (D) The binding of HOXB13 to VGLL4 promoter regions (S1/S2). (E) The interaction between HOXB13 and VGLL4 was determined by luciferase reporter gene assay. ***P<0.001 vs. Ov-NC. (F) The direct binding of HOXB13 and VGLL4 promoter was confirmed using chromatin immunoprecipitation. ***P<0.001 vs. IgG. (G) Protein expression levels of downstream effectors of the Hippo signaling pathway, including CCN2, Cyr61 and AREG, were determined with western blot analysis and semi-quantified. **P<0.01, ***P<0.001 vs. Control; # P<0.05, ### P<0.001 vs. Ov-HOXB13 + si-NC. AREG, amphiregulin; CCN2, cellular communication network factor 2; Cyr61, cysteine rich angiogenic inducer 61; HOXB13, homeobox B13; F-luc/R-Luc, Firefly luciferase/ Renilla luciferase; MUT, mutant-type; NC, negative control; Ov, overexpression; si, small interfering RNA; TEAD4, TEA domain 4; TSS, transcription start site; VGLL4, vestigial-like family member 4; WT, wild-type; RT-qPCR, reverse transcription-quantitative PCR; S1, site 1; S2, site 2.

Article Snippet: The membranes were then incubated with following primary antibodies overnight at 4°C: Anti-HOXB13 (1:1,000; cat. no. ab201682; Abcam), anti-proliferating cell nuclear antigen (PCNA; 1:1,000; cat. no. ab92552; Abcam), anti-Ki-67 (1:1,000; cat. no. ab92742; Abcam), anti-MMP2 (1:1,000; cat. no. ab92536; Abcam), anti-MMP9 (1:1,000; cat. no. ab76003; Abcam), anti-issue inhibitor of metalloproteinases 1 (TIMP-1; 1:1,000; cat. no. ab211926; Abcam), anti-Bcl-2 (1:1,000; cat. no. ab182858; Abcam), anti-Bax (1:1,000; cat. no. ab32503; Abcam), anti-cleaved caspase-3 (1:500; cat. no. ab32042; Abcam), anti-caspase-3 (1:1,000; cat. no. ab32351; Abcam), anti-TEAD4 (1:1,000; cat. no. ab155244; Abcam), anti-cellular communication network factor 2 (CCN2; 1:1,000; cat. no. ab209780; Abcam), anti-cysteine rich angiogenic inducer 61 (Cyr61; 1:1,000; cat. no. ab230947; Abcam), anti-amphiregulin (AREG; 1:1,000; cat. no. ab180722; Abcam) and anti-GAPDH (1:2,000; cat. no. ab181602; Abcam).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot, Binding Assay, Luciferase, Reporter Gene Assay, Chromatin Immunoprecipitation, Mutagenesis, Negative Control, Over Expression, Small Interfering RNA, Real-time Polymerase Chain Reaction

Journal: Drug Metabolism and Disposition

Article Title: Dexamethasone-Induced Liver Enlargement Is Related to PXR/YAP Activation and Lipid Accumulation but Not Hepatocyte Proliferation

doi: 10.1124/dmd.120.000061

Figure Lengend Snippet: Effect of Dex on YAPsignaling pathway. (A and B) Western blot analysis and quantification of total YAP, nuclear YAP and cytoplasmic phosphorylated YAP protein of liver samples after a 5-day treatment with the vehicle or Dex. (C and D) Western blot analysis and quantification of YAP downstream proteins of liver samples after the vehicle or Dex treatment. Data are expressed as mean ± S.D. (n = 3). *P < 0.05; **P < 0.01 compared with the vehicle group. (E) Confocal microscopy displaying PXR and YAP distribution in HepG2 cells treated with DMSO or 100 μM of Dex for 6 hours. Scale bar, 40 μm. (F) Quantification of immunofluorescence double staining of YAP and PXR. Data are expressed as mean ± S.D. (n = 3). *P < 0.05 compared with the control group. ANKRD1, ankyrin repeat domain 1; CTGF, connective tissue growth factor; CYR61, cysteine-rich angiogenic inducer 61; DAPI, 4′,6-diamidino-2-phenylindole; p-YAP, phosphorylated YAP.

Article Snippet: Rabbit polyclonal anti-CCNA1 (Cat. D220507), anti–phosphorylated YAP (Cat. D151452), anti–ankyrin repeat domain 1 (Cat. D121628), anti-cysteine-rich angiogenic inducer 61 (Cat. D122190), anti–connective tissue growth factor (Cat. D160212), anti–epidermal growth factor receptor (EGFR) (Cat. D260292), anti–mesenchymal-epithelial transition factor (MET) (Cat. D160981), and anti–lamin B1 (Cat. D220926) antibodies were acquired from Sangon Biotechnology (Sangon Tech, Shanghai, China).

Techniques: Western Blot, Confocal Microscopy, Immunofluorescence, Double Staining

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