Journal: Oncology Reports

Article Title: Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation

doi: 10.3892/or.2026.9086

Figure Lengend Snippet: Knockdown of CCT2 inhibits STAT3 signaling activation in hepatocellular carcinoma cells. The protein levels of STAT3, p-STAT3, MCL1, MMP2 and SOX2 in (A) Huh-7 and (B) HCCLM3 cells were measured by western blotting. The protein levels of (C) p-STAT3, (D) MCL1, (E) MMP2 and (F) SOX2 in subcutaneous tumor tissue were detected by immunohistochemical staining. *P<0.05, **P<0.01 vs. sh-NC. CCT2, chaperonin containing TCP1 subunit 2; sh, short hairpin; NC, negative control; p, phosphorylated; MCL1, myeloid cell leukemia sequence 1; SOX2, SRY-box transcription factor 2.

Article Snippet: The primary antibodies were as follows: CCT2 (cat. no. 24896-1-AP), β-actin (cat. no. 66009-1-Ig), MMP2 (cat. no. 10373-2-AP), myeloid cell leukemia sequence 1 (MCL1; cat. no. 16225-1-AP) and SRY-box transcription factor 2 (SOX2; cat. no. 11064-1-AP; all Proteintech Group, Inc.) and STAT3 (cat. no. 4904) and phosphorylated (p-)STAT3 (Tyr705; cat. no. 4113; both Cell Signaling Technology, Inc.) The membranes were washed three times in TBST (0.1% Tween-20) for 5 min each at room temperature.

Techniques: Knockdown, Activation Assay, Western Blot, Immunohistochemical staining, Staining, Negative Control, Sequencing

Journal: Cell Death & Disease

Article Title: O-GlcNAcylation of YAP1 promotes lung transplant ischemia-reperfusion injury via binding to HIF1α transcription factor and activating autophagy and mitophagy

doi: 10.1038/s41419-026-08548-w

Figure Lengend Snippet: A Western blot analysis revealed that hypoxia-reoxygenation (0, 2, 4, 6, 12 h) treatment gradually increased the O-GlcNAc modification level of total cellular proteins and the expression of the key modifying enzyme OGT. B Western blot analysis showed that ischemia-reperfusion (2, 6, 12 h) gradually increased the O-GlcNAc modification level of total lung tissue proteins and the expression of OGT. C YAP1 protein immunoprecipitation experiments demonstrated that hypoxia-reoxygenation (0, 2, 4, 6, 12 h) treatment gradually increased the O-GlcNAc modification level of cellular YAP1 protein. D YAP1 protein immunoprecipitation experiments revealed that ischemia-reperfusion (2, 6, 12 h) gradually increased the O-GlcNAc modification level of YAP1 protein in lung tissue. E Flag-YAP1 protein immunoprecipitation with M2 Magnetic Beads experiments revealed that overexpression of wild-type OGT significantly increased the O-GlcNAc modification level of wild-type Flag-YAP1 protein and its binding to the HIF1A transcription factor in alveolar epithelial cells under hypoxia-reoxygenation (12 h). Meanwhile, overexpression of OGT with a serine mutation at the O-GlcNAc modification site had no such effect. Additionally, OGT knockdown significantly inhibited the O-GlcNAc modification level of wild-type Flag-YAP1 protein and its binding to the HIF1A transcription factor in alveolar epithelial cells under hypoxia-reoxygenation (12 h). F Chromatin immunoprecipitation (ChIP) experiments for HIF1α found that overexpression of wild-type OGT significantly increased the binding level of the HIF1A transcription factor to the promoters of autophagy genes ( HMGB1 , DAPK , and LC3-II ) and mitophagy genes ( FUNDC1 , PINK1 , TBK1 ) in alveolar epithelial cells under hypoxia-reoxygenation (12 h). In contrast, overexpression of OGT with a serine mutation at the O-GlcNAc modification site had no such effect. Moreover, OGT knockdown significantly inhibited the binding level of the HIF1A transcription factor to the promoters of these autophagy and mitophagy genes in alveolar epithelial cells under hypoxia-reoxygenation (12 h). Data represent mean ± SEM ( N = 3), * P < 0.05, ** P < 0.01, *** P < 0.001, compared with indicated group by two-way ANOVA along with Tukey’s post hoc test. G Dual luciferase reporter experiments demonstrated that overexpression of wild-type OGT significantly increased the promoter activity of autophagy genes ( HMGB1 , DAPK , and LC3-II ) and mitophagy genes ( FUNDC1 , PINK1 , TBK1 ) in alveolar epithelial cells under hypoxia-reoxygenation (12 h). Conversely, overexpression of OGT with a serine mutation at the O-GlcNAc modification site had no such effect. Additionally, OGT knockdown significantly inhibited the promoter activity of these autophagy and mitophagy genes in alveolar epithelial cells under hypoxia-reoxygenation (12 h). Data represent mean ± SEM ( N = 3), * P < 0.05, ** P < 0.01, *** P < 0.001, compared with indicated group by two-way ANOVA along with Tukey’s post hoc test.

Article Snippet: The fragmented chromatin was incubated with an antibody specific to the HIF1A transcription factor (Cat No. 20960-1-AP, 1:200, Proteintech, Wuhan, China).

Techniques: Western Blot, Modification, Expressing, Immunoprecipitation, Magnetic Beads, Over Expression, Binding Assay, Mutagenesis, Knockdown, Chromatin Immunoprecipitation, Luciferase, Activity Assay