Journal: Journal of Biological Chemistry

Article Title: Selective Enhancing Effect of Early Mitotic Inhibitor 1 (Emi1) Depletion on the Sensitivity of Doxorubicin or X-ray Treatment in Human Cancer Cells

doi: 10.1074/jbc.m112.446351

Figure Lengend Snippet: FIGURE 7. Synergistic anticancer effect of Emi1 knockdown in combination with doxorubucin in normal cells. A, Emi1 or control siRNA was transfected into NHDF cells, and cells were collected after 48 h. The indicated proteins in siRNA-treated NHDF cells were examined by Western blotting. B, Emi1 siRNA and/or cyclin A siRNA were transfected into NHDF cells. The left panel shows the expression of Emi1 and cyclin A examined by Western blot analysis after 48 h of siRNA transfection. -Actin expression was used as a loading control. The right panel shows DAPI staining and percentage of cells with polyploidy after Emi1 siRNA and/or cyclin A siRNA transfection. Cells were stained with DAPI to visualize the nuclei, and percentage of cells with polyploidy was determined by DNA content analysis after PI staining using a flow cytometer. *, p 0.05. C, flow cytometric analysis of annexin V and PI staining in control and Emi1 siRNA treated NHDF or HFL III cells after treatment with doxorubucin (DOXY; 0.5 g/ml) for 12 h. We performed three independent experiments.

Article Snippet: Human cyclin A siRNA (sc-29282) and human E2F1 siRNA (sc-61861) were obtained from Santa Cruz Biotechnology.

Techniques: Knockdown, Control, Transfection, Western Blot, Expressing, Staining, Flow Cytometry

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Super enhancer-driven LINC01013 mediates hypoxia-induced mitochondrial dysfunction by HSPA9 to determine pulmonary arterial smooth muscle cell fate

doi: 10.1007/s00018-025-06071-3

Figure Lengend Snippet: Silencing of LINC01013 inhibited hypoxia-induced hPASMC proliferation and inflammation. A , B hPASMC proliferation was determined by CCK8 and EdU incorporation assays ( n = 6). EdU (red), DAPI (blue), scale bar, 50 μm. C Western blotting analysis of PCNA, cyclin A and cyclin D in hPASMCs ( n = 6). D RT‒qPCR analysis showed the mRNA levels of TNF-α and IL-6 after LINC01013 knockdown, β-actin was used as a internal reference gene ( n = 6). E Western blotting analysis of TNF-α and IL-6 in hPASMCs. ( n = 6). All values are presented as the mean ± SEM. Statistical analysis was performed with one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001. Nor, normoxia; Hyp, hypoxia; NC, negative control; si, siRNA

Article Snippet: The antibody against CEBPB (PB9171, BA0670, 1:500, Boster, Wuhan, China), H3K27ac (A7253, 1:500, ABclonal, Wuhan, China), H3K4me1 (A2355, 1:500, Wuhan, China), PCNA (A00125, 1:500, Boster, Wuhan, China), Cyclin A (PB0515, 1:500, Boster, Wuhan, China), Cyclin D (BM4272, 1:500, Boster, Wuhan, China), IL-6 (AF7236, 1:500, Beyotime, Shanghai, China), TNF-α (AF8208, 1:500, Beyotime, Shanghai, China), PKM2 (4053, 1:1000, Cell Signaling, MA, US), HK II (66974-1-Ig, 1:1000, Proteintech, IL, USA), PDH (2784, 1:1000, Cell Signaling, MA, US), HSPA9 (14887-1-AP, 1:5000, Proteintech, IL, USA), VDAC1 (10866-1-AP, 1:5000, Proteintech, IL, USA), and β-actin (TA-09, 1:1000, ZSGB‐BIO, Beijing, China) was incubated at 4 °C overnight, followed by incubation with appropriate horseradish peroxidase-conjugated secondary antibodies at room temperature for 1 h, and proteins were visualized with enhanced chemiluminescence reagents.

Techniques: Western Blot, Knockdown, Negative Control