Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Inhibition of the interaction between Hippo/YAP and Akt signaling with ursolic acid and 3′3-diindolylmethane suppresses esophageal cancer tumorigenesis

doi: 10.4196/kjpp.2023.27.5.493

Figure Lengend Snippet: After TE-8 and TE-12 cells were treated with UA (20 μM), DIM (50 μM), and a combination for 48 h. Akt/p-Akt and Gsk-3β/p-GSK 3β protein levels were examined by Western blot analysis. The quantification of protein bands was estimated by ImageJ software. GAPDH was used as the internal control. Data are expressed as the mean ± standard error of the mean. CONT, control; ESCC, esophageal squamous cell carcinoma; n.s., no significant. *, compared to the control; #, compare with UA plus DIM combination treatment. * or #, p < 0.05; ** or ##, p < 0.01.

Article Snippet: Primary antibodies against YAP, Mst1, Mst2, caspase-9, c-caspase-9, p-YAP, PARP, c-PARP, Akt, p-Akt, Mob-1, p-Mob1, p-GSK-3β, E-cadherin, MMP-9, CDK4, CDK6, cyclin D1, Sav1, p-PTEN, PTEN, and GAPDH were obtained from Cell Signaling Technology, and primary antibodies against MMP-13, Rassf1, and CTGF were purchased from Santa Cruz Biotechnology Inc. UA was obtained from Cayman Chemical Company, and DIM was purchased from LKT Laboratories.

Techniques: Western Blot, Software

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Inhibition of the interaction between Hippo/YAP and Akt signaling with ursolic acid and 3′3-diindolylmethane suppresses esophageal cancer tumorigenesis

doi: 10.4196/kjpp.2023.27.5.493

Figure Lengend Snippet: Ursolic acid (UA) plus 3,3’-diindolylmethane (DIM) combination treatment can induce apoptosis by promoting cleaved-PARP and caspase-9, increasing the G1 phase by upregulating CDK4, CDK6, and Cyclin D1 protein expression, and inhibiting migration by regulating E-cadherin, MMP-9, and MMP-13 in esophageal squamous cell carcinoma (ESCC). UA plus DIM combination treatment lowered p-Akt and p-Gsk-3β expression in ESCC. UA plus DIM combination activated the Hippo signaling pathway leading to the inhibition of YAP de-phosphorylation and nuclear translocation and suppression of its downstream gene CTGF further restrained cell growth in ESCC. UA and DIM combination treatment induced the Hippo pathway activation by inhibiting the PI3K/Akt pathway in ESCC. UA plus DIM combination treatment can induce YAP inhibition by conversely inhibiting the PI3K/Akt pathway through up-regulated PTEN. UA plus DIM stimulated a feedback loop between the PI3K/Akt signaling pathway and Hippo signaling pathways in ESCC.

Article Snippet: Primary antibodies against YAP, Mst1, Mst2, caspase-9, c-caspase-9, p-YAP, PARP, c-PARP, Akt, p-Akt, Mob-1, p-Mob1, p-GSK-3β, E-cadherin, MMP-9, CDK4, CDK6, cyclin D1, Sav1, p-PTEN, PTEN, and GAPDH were obtained from Cell Signaling Technology, and primary antibodies against MMP-13, Rassf1, and CTGF were purchased from Santa Cruz Biotechnology Inc. UA was obtained from Cayman Chemical Company, and DIM was purchased from LKT Laboratories.

Techniques: Expressing, Migration, Inhibition, De-Phosphorylation Assay, Translocation Assay, Activation Assay

Journal: Cell Death Discovery

Article Title: Suppressed Akt/GSK-3β/β-catenin signaling contributes to excessive adipogenesis of fibro-adipogenic progenitors after rotator cuff tears

doi: 10.1038/s41420-023-01618-4

Figure Lengend Snippet: a , b The oil red staining and statistical analysis of RCT-FAPs (FAPs collected from supraspinatus muscle one week after tendon tears) with different treatments. Freshly isolated RCT-FAPs were cultured in AIM, AIM with Akt agonist SC-79, AIM with SC-79 and β-catenin inhibitor KYA1797K for 7 days ( n = 3). Scale bar = 100 µm. AIM adipogenic induction medium. SC-79, Akt signaling agonist; KY1797K, β-catenin inhibitor. c Relative mRNA expression level of adipogenic-related genes for RCT-FAPs after adipogenic differentiation with different treatments ( n = 3). d Protein level of β-catenin, p-Akt, t-Akt, PPARγ, p-GSK-3β, GSK-3β, and GAPDH for RCT-FAPs treated by AIM, AIM with SC-79, AIM with SC-79 and KYA1797K. e The scheme of Akt/GSK3β/β-catenin signaling, which regulated adipogenesis of FAPs. f Relative mRNA expression level of β-catenin after transfected with β-catenin siRNA in RCT-FAPs ( n = 3). g The protein levels of β-catenin and PPARγ after transfected with β-catenin siRNA in RCT-FAPs ( n = 3). h , i The oil red staining and statistical analysis of RCT-FAPs after β-catenin siRNA transfection and adipogenic induction ( n = 3). Scale bar = 100 µm. j Relative mRNA expression level of adipogenic genes for RCT-FAPs after β-catenin siRNA transfection and adipogenic induction. ( n = 3). k Protein level of β-catenin, p-Akt, t-Akt, PPAR γ, p-GSK-3β, GSK-3β, and GAPDH for RCT-FAPs and ATT-FAPs. * indicated p < 0.05, ** indicated p < 0.01, and *** indicated p < 0.001.

Article Snippet: Primary antibodies were incubated overnight at 4 °C: anti-GAPDH (CST, cat#2118 L), anti-Akt (CST, cat#4865), anti-phospho-Akt (CST, cat#4060), anti-GSK-3β (Santa Cruz, cat#sc-53931), anti-phospho-GSK-3β (CST, cat#93336), anti-β-catenin (CST, cat#8480), anti-PPARγ (CST, cat#2435).

Techniques: Staining, Isolation, Cell Culture, Expressing, Transfection