Phosphorylation analysis of the Hippo-YAP pathway using Phos-tag.

Phosphorylation is an essential regulatory mechanism in cells that modifies diverse substrates, such as proteins, carbohydrates, lipids, and nucleotides. Protein phosphorylation regulates function, subcellular localization, and protein-protein interactions. Protein kinases and phosphatases catalyze this reversible mechanism, subsequently influencing signal transduction. The dysregulation of protein phosphorylation leads to many diseases, such as cancer, neurodegenerative diseases, and metabolic diseases. Therefore, analyzing the phosphorylation status and identifying protein phosphorylation sites are critical for elucidating the biological functions of specific phosphorylation events. Unraveling the critical phosphorylation events associated with diseases and specific signaling pathways is promising for drug discovery. To date, highly accurate and sensitive approaches have been developed to detect the phosphorylation status of proteins. In this review, we discuss the application of Phos-tag to elucidate the biological functions of Hippo pathway components, with emphasis on the identification and quantitation of protein phosphorylation under physiological and pathological conditions. SIGNIFICANCE: We here provide a comprehensive overview of Phos-tag technique-based strategies to identify phosphorylated proteins at the cellular level in the Hippo-YAP pathway that comprises a major driving force for cellular homeostasis. We clarify the links of applying Phos-tag in elucidating the biological functions of the Hippo pathway components with particular attention to the identification and quantitation of protein phosphorylation under physiological and pathological conditions. We believe that our paper will make a significant contribution to the literature because these detailed phosphorylation modifications and functional diversity of the Hippo pathway components in physiological and pathological processes are only beginning to come to the fore, highlighting the potential for discovering new therapeutic targets. Moreover, this line of research can provide further insight into the inextricable link between phos-tag applications as a molecular tool and cellular signaling modality, offering new directions for an integrated research program toward understanding cellular regulation at the molecular level. Given the broad research and practical applications, we believe that this paper will be of interest to the readership of your journal.

Extracts of Perilla frutescens var. Acuta (Odash.) Kudo Leaves Have Antitumor Effects on Breast Cancer Cells by Suppressing YAP Activity.

Yes-associated protein (YAP)/WW domain-containing transcription factor (TAZ) is critical for cell proliferation, survival, and self-renewal. It has been shown to play a crucial oncogenic role in many different types of tumors. In this study, we investigated the antitumor effect of the extracts of Perilla frutescens var. acuta (Odash.) Kudo leaves (PLE) on Hippo-YAP/TAZ signaling. PLE induced the phosphorylation of YAP/TAZ, thereby inhibiting their activity. In addition, the treatment suppresses YAP/TAZ transcriptional activity via the dissociation of the YAP/TAZ-TEAD complex. To elucidate the molecular mechanism of PLE in the regulation of YAP activity, we treated WT and cell lines with gene knockout (KO) for Hippo pathway components with PLE. The inhibitory effects of PLE on YAP-TEAD target genes were significantly attenuated in LATS1/2 KO cells. Moreover, we found the antitumor effect of PLE on MDA-MB-231 and BT549, both of which are triple-negative breast cancer (TNBC) cell lines. PLE reduced the viability of TNBC cells in a dose-dependent manner and induced cell apoptosis. Further, PLE inhibited the migration ability in MDA-MB-231 cells. This ability was weakened in YAP and TEAD-activated clones suggesting that the inhibition of migration by PLE is mainly achieved by regulating YAP activity. Taken together, the results of this study indicate that PLE suppressed cell growth and increased the apoptosis of breast cancer (BC) cells via inactivation of YAP activity in a LATS1/2-dependent manner.

Stretchable Metamaterial Absorber Using Liquid Metal-Filled Polydimethylsiloxane (PDMS).

A stretchable metamaterial absorber is proposed in this study. The stretchability was achieved by liquid metal and polydimethylsiloxane (PDMS). To inject liquid metal, microfluidic channels were fabricated using PDMS powers and microfluidic-channel frames, which were built using a three-dimensional printer. A top conductive pattern and ground plane were designed after considering the easy injection of liquid metal. The proposed metamaterial absorber comprises three layers of PDMS substrate. The top layer is for the top conductive pattern, and the bottom layer is for the meandered ground plane. Flat PDMS layers were inserted between the top and bottom PDMS layers. The measured absorptivity of the fabricated absorber was 97.8% at 18.5 GHz, and the absorption frequency increased from 18.5 to 18.65 GHz as the absorber was stretched from its original length (5.2 cm) to 6.4 cm.

Microfluidic tunable inkjet-printed metamaterial absorber on paper.

In this paper, we propose a novel microfluidic tunable metamaterial (MM) absorber printed on a paper substrate in silver nanoparticle ink. The metamaterial is designed using a periodic array consisting of square patches. The conductive patterns are inkjet-printed on paper using silver nanoparticle inks. The microfluidic channels are laser-etched on polymethyl methacrylate (PMMA). The conductive patterns on paper and the microfluidic channels on PMMA are bonded by an SU-8 layer that is also inkjet-printed on the conductive patterns. The proposed MM absorber provides frequency-tuning capability for different fluids in the microfluidic channels. We performed full-wave simulations and measurements that confirmed that the resonant frequency decreased from 4.42 GHz to 3.97 GHz after the injection of distilled water into the microfluidic channels. For both empty and water-filled channels, the absorptivity is higher than 90% at horizontal and vertical polarizations.

Flexible inkjet-printed metamaterial absorber for coating a cylindrical object.

In this paper, a novel flexible inkjet-printed metamaterial absorber is proposed. The unit cell of the metamaterial is designed with a modified Jerusalem-cross ring resonator and is inkjet printed with silver nanoparticle ink on a flexible polymer film. All fabrication processes are performed using a commercial printer (EPSON WF-7011). The absorber's flexibility and absorption performance are demonstrated by measuring the absorption ratio after coating the proposed absorber on a cylindrical object with a radius of 4.56 cm. An absorption rate exceeding 99% is achieved at 9.21 GHz for both flat and cylindrical surfaces. In addition, the cylindrical model attains an absorption rate higher than 96% for all polarization angles, and a high absorption rate of 95% is preserved until the incident angle is less than 30þ.