viously described [52]. two.five. Western Blot Evaluation On day 8 of cell differentiation, whole cell protein lysates from differentiated cells had been ready, resolved by ten sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and transferred to nitrocellulose membranes. Target proteins, such as phospho-protein kinase B (P-Akt), Akt, phospho-extracellular signal-regulated kinase (P-ERK), ERK, phospho-cJun N-terminal kinase (P-JNK), JNK, phospho-P38 (P-P38), P38, peroxisome proliferatoractivated receptor gamma (PPAR-), CCAAT/enhancer-binding protein alpha (C/EBP-), C/EBP-, glucocorticoid receptor (GR), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH), have been detected making use of principal antibodies and horseradish peroxidase-labeled anti-rabbit secondary antibodies (Cell Signaling Technology, Danvers, MA, USA). Target proteins had been visualized applying ECL Plus Western blotting detection reagents (GE Healthcare, Piscataway, NJ, USA). Protein levels were determined densitometrically applying a chemiluminescence method (FUSION Solo, PEQLAB Biotechnologie GmbH, Erlangen, Germany), as previously described [53]. 2.6. Statistical Analysis Statistical significance was determined making use of one-way analysis of variance and multiple comparisons with Bonferroni correction. Statistical significance was set at p 0.05. All analyses were performed working with SPSS Statistics ver. 19.0 (SPSS Inc., Chicago, IL, USA).2.six. Statistical AnalysisBiomolecules 2021, 11,Statistical significance was determined utilizing one-way analysis of variance and several comparisons with Bonferroni correction. Statistical significance was set at p 0.05.21 5 of All analyses have been performed utilizing SPSS Statistics ver. 19.0 (SPSS Inc., Chicago, IL, USA). three. Final results three. Outcomes three.1. Network Pharmacology Evaluation 3.1. Network Pharmacology Evaluation three.1.1. Target Prediction and Screening of HDAC5 Inhibitor web Possible Targets 3.1.1. Target Prediction and Screening of Potential Targets The SwissTargetPrediction database was made use of to predict the targets of hispidulin as well as the SwissTargetPrediction database was made use of to predict the targets of hispidulin p-synephrine. In data preprocessing, 103 and 32 verified targets of hispidulin and and and p-synephrine. In data preprocessing, 103 and 32 verified targets of hispidulin psynephrine, respectively, were screened. Additionally, 94899489 obesity-related targets were p-synephrine, respectively, were screened. Moreover, obesity-related targets had been acquired from the HDAC6 Inhibitor site GeneCards database, and also the relevance score was utilised as a cut-off worth. acquired from the GeneCards database, as well as the relevance score was made use of as a cut-off Based on the relevance score, 1897 obesity-related targets belonging for the leading the leading 20 worth. Determined by the relevance score, 1897 obesity-related targets belonging to 20 have been used for thefor the analysis. As shown in Figure 1, the predicted targets of hispidulin and were applied analysis. As shown in Figure 1, the predicted targets of hispidulin and psynephrine shared 53 and 23 targets, respectively, with obesity-related targets. Therefore, these p-synephrine shared 53 and 23 targets, respectively, with obesity-related targets. As a result, targets targets were selected as prospective targets (Tables2).and two). these have been chosen as prospective targets (Tables 1 andFigure Venn diagrams of predicted targets of compounds and obesity-related targets. (A) Venn Figure 1.1. Venndiagrams of predicted targets of compounds and obesity-related targets. (A) Venn diagram of hispidulin-predi