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

Journal: The Journal of Clinical Investigation

Article Title: Wilms tumor 1 impairs apoptotic clearance of fibroblasts in distal fibrotic lung lesions

doi: 10.1172/JCI188819

Figure Lengend Snippet: ( A ) Schematic representation of the animal experiment involving PDGFR α CreERT (control) and PDGFR α CreERT WT1 OE (cWT1 OE ) mice. Mice were treated repeatedly with bleomycin via intratracheal administration and with tamoxifen via intraperitoneal injection. ( B ) Quantification of Wt1 gene transcripts by RT-PCR in total lung RNA isolated from control and cWT1 OE mice ( n = 6/group). ** P < 0.01, by 2-tailed Student’s t test. ( C ) Representative confocal images of lung sections from control and cWT1 OE mice co-immunostained for WT1 (red), vimentin (green), and DAPI (blue). Scale bars: 20 μm. ( D ) Representative images of Masson’s trichrome–stained lung sections from control and cWT1 OE mice. Original magnification, ×4 and ×20, respectively. Scale bars: 1,500 μm and 200 μm, respectively. ( E ) The percentage of fibrotic area was quantified in control and cWT1 OE mice using BZ-X image analysis ( n = 6/group). **** P < 0.0001, by 2-tailed Student’s t test. ( F ) Hydroxyproline levels were measured in the right lungs of control and cWT1 OE mice ( n = 6/group). *** P < 0.001, by 2-tailed Student’s t test. ( G ) Lung resistance was assessed using FlexiVent in control and cWT1 OE mice treated with bleomycin ( n = 6/group). * P < 0.01, by 2-tailed Student’s t test. ( H ) Proliferation of fibroblasts was quantified using a BrdU incorporation assay in lung cultures from control and cWT1 OE mice treated with bleomycin ( n = 3/group). * P < 0.05, by 2-tailed Student’s t test. ( I ) Fibroblasts from the lung cultures of control and cWT1 OE mice treated with bleomycin were treated with anti-Fas antibody for 24 hours, followed by TUNEL staining (red). Representative confocal images are shown; nuclei are stained with DAPI (blue). Original magnification, ×20. Scale bars: 100 μm. ( J ) Percentage of TUNEL + fibroblasts in total DAPI + fibroblasts ( n = 3/group). * P < 0.05 and ** P < 0.01, by 1-way ANOVA. Data are representative of 2 independent experiments with similar findings.

Article Snippet: Cell proliferation was evaluated using the BrdU kit (Cell Signaling Technology) as described previously ( ).

Techniques: Control, Injection, Reverse Transcription Polymerase Chain Reaction, Isolation, Staining, BrdU Incorporation Assay, TUNEL Assay