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Krüppel-like factor 7 (KLF7) is a negative regulator of preadipocyte differentiation. Our previous KLF7 ChIP-seq analysis showed that the binding motif of PU.1 was found among the KLF7 binding peaks, indicating that an interaction between KLF7 and PU.1 at preadipocyte gene promoters and other regulatory elements might be common. Here, Co-IP and FRET assays are used to confirm that PU.1 can directly bind to KLF7 and enhance the transcription activity of cyclin-dependent kinase inhibitor 3 ( CDKN3), which is a downstream target gene of KLF7. We show that the PU.1 expression level is decreased during preadipocyte differentiation. Furthermore, PU.1 overexpression and knockdown experiments reveal that PU.1 negatively regulates chicken preadipocyte differentiation, as evidenced by appropriate changes in lipid droplet accumulation and altered expressions of PPARγ, FAS, and PLIN. In addition, PU.1 overexpression promotes preadipocyte proliferation, while knockdown of PU. 1 inhibits preadipocyte proliferation. We further demonstrate that PU.1 inhibits differentiation and promotes proliferation in preadipocytes, in part by directly interacting with KLF7.

Krüppel-like transcription factor 7 (KLF7) promotes preadipocyte proliferation; however, its target gene in this process has not yet been identified. Using KLF7 ChIP-seq analysis, we previously showed that a KLF7-binding peak is present upstream of the cyclin-dependent kinase inhibitor 3 gene ( CDKN3) in chicken preadipocytes. In the present study, we identify CDKN3 as a target gene of KLF7 that mediates the effects of KLF7 on preadipocyte proliferation. Furthermore, 5′-truncating mutation analysis shows that the minimal promoter is located between nt –160 and nt –7 (relative to the translation initiation codon ATG) of CDKN3. KLF7 overexpression increases CDKN3 promoter activity in the DF-1 and immortalized chicken preadipocyte (ICP1) cell lines. Deletion of the putative binding site of KLF7 abolishes the promotive effect of KLF7 overexpression on CDKN3 promoter activity. Moreover, CDKN3 knockdown and overexpression assays reveal that CDKN3 enhances ICP1 cell proliferation. Flow cytometry analysis shows that CDKN3 accelerates the G1/S transition. Furthermore, we find that KLF7 promotes ICP1 cell proliferation via Akt phosphorylation by regulating CDKN3. Taken together, our results suggest that KLF7 promotes preadipocyte proliferation by activating the Akt signaling pathway by cis-regulating CDKN3, thus driving the G1/S transition.

Intentional islanding restoration of distribution systems with multi-generations is of great importance to ensure the power supply of critical loads under extreme conditions, which is beneficial to improve the reliability of distribution systems. There are transient voltage and frequency fluctuations in the process of intentional islanding restoration, when the loads and distributed generations are gradually connected to the grid. The safety and stability of the intentional islanding are affected by the fluctuations, and networking process may fail in serious cases. Hence, the rapid power response of the energy storage system is utilized to suppress voltage and frequency fluctuations. A fluctuation suppression model based on energy storage system control is established, where a voltage and current double-loop feed-forward disturbance compensation control system is designed. A vector control method for energy storage system with improved dual-loop control is proposed, which solves the problems of traditional V/f control voltage offset and excessive voltage fluctuation. MATLAB/Simulink is used to build simulation models in different control modes in accordance with the black-start principle. The simulation results show that the improved double-loop control based on the vector method has a stronger anti-interference ability and significantly improved the islanding black-start self-organizing networking process. Voltage and frequency fluctuations are reduced, and the dynamic response performance of the system is improved.