Fig. 6 | Signal Transduction and Targeted Therapy

Fig. 6

From: LncRNA Wee1-AS coordinates oxidative fatty acid metabolism through the activation of mitochondrial CDK1/CYCLIN B1

Fig. 6The alternative text for this image may have been generated using AI.

Wee1-AS enhances mitochondrial homeostasis via activation of CDK1/CYCLIN B1. a Primary hepatocytes were treated with 25 μM palmitic acid (PA)-conjugated with 0.05% bovine serum albumin (BSA) for the indicated times. The mitochondrial fraction was prepared and analyzed via western blotting. The values are represented the means ± SDs (n = 3). The data were analyzed via two-way ANOVA. *P < 0.05 vs BSA treatment for 1 h. b, c Primary hepatocytes were transfected with ASO-control (ctl) or ASO-Wee1-AS. The values are presented as the means ± SDs (n = 4). The data were analyzed via the Mann‒Whitney test. b The mitochondrial fraction was prepared and analyzed by western blotting. The intensity of each protein band was quantified via ImageJ and normalized to that of COX4 or CDK1. *P < 0.05 vs ASO-control (ctl). c Cells were stained with 100 nM MitoTracker Red CMXRos (red). Immunostaining was performed for CYCLIN B1 (green). Scale bar, 20 μm. The fluorescence intensity in at least 100 cells was quantified via Image J software. The percentage of co-localization was determined via the JACoP plugin of ImageJ software. The amount of mitochondrial CYCLIN B1 was normalized to the intensity of MitoTracker signal. ***P < 0.001 vs ASO-control (ctl). d, e Tissue extracts were prepared from the livers of the mice shown in Fig. 2a. The values are presented as the means ± SDs (n = 4). The data were analyzed via the Mann‒Whitney test. d The mitochondrial fraction was prepared and analyzed via western blotting. *P < 0.05 vs HFD with AAV-GFP. e The mitochondrial fraction was prepared and the phosphorylation of the indicated proteins was analyzed via immunoprecipitation (IP) with an anti-phospho-serine/threonine antibody and probed with specific antibodies. The lanes were run on the same gel but were noncontiguous. The intensity of each protein band was quantified via ImageJ. *P < 0.05 vs HFD with AAV-GFP. f Primary hepatocytes were transfected with si-GFP/si-Cdk1 or pcDNA-empty vector (Ctl)/pcDNA-Wee1-AS (Wee1-AS), and then treated with free fatty acids (200 μM oleic acid/100 μM palmitic acid) for 24 h. Lipid accumulation was assessed via BODIPY staining. Scale bar, 20 μm. The relative fluorescence area in at least 100 cells was quantified via Image J software. The values are presented as the means ± SDs (n = 10). The data were analyzed via two-way ANOVA. ***P < 0.001 vs pcDNA-empty vector (Ctl) with si-GFP; ###P < 0.001 vs pcDNA-Wee1-AS (Wee1-AS) with si-GFP

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