cyclin d 3  (Cell Signaling Technology Inc)

Journal: MedComm

Article Title: Inhibition of FOS‐Like Antigen 1 Reduces Chemoresistance to Temozolomide Through Stemness Reprogramming via IL‐6/STAT3 Tyr705 Pathway

doi: 10.1002/mco2.70593

Figure Lengend Snippet: Inhibition of FOSL1 in GBM cells reverses TMZ response. (A, B) Representative histogram of protein expression, including FOSL1 and cell cycle‐associated molecules, in GBM patient's cells was shown (top panel). Protein expression in patients with GBM‐derived cells ( n = 6), including FOSL1 and cell cycle‐associated molecules, was analyzed by flow cytometry and quantified using FlowJo V10 (bottom panel). (C) FOSL1 mRNA expression (left panel) in U87MG treated with si‐control or si‐FOSL1 analyzed using qPCR. GAPDH was used as a control gene for relative quantification. FOSL1 protein expression in U87MG treated with si‐control or si‐FOSL1 was analyzed by flow cytometry and quantified using FlowJo V10 (right panel). (D) Expression of G0/G1 to S phase transition‐related proteins, including CDK4, cyclin D, CDK2, and cyclin E, analyzed using flow cytometry and quantified using FlowJo V10. E Population of the G0/G1 phase of U87MG cells stained with CCS1 and analyzed using flow cytometry. Data were quantified using FlowJo V10. (F) Representative images of colony formation showing viable U87MG cells treated with si‐control, si‐FOSL1 or TMZ (purple). (G) Proliferation of U87MG cells calculated using the WST‐8 reduction assay following the manufacturer's instructions. *p < 0.05; **p < 0.005; ***p < 0.0005; paired t ‐test ( n = 3).

Article Snippet: For intracellular protein analysis, U87MG cells and patient‐derived GBM cells were collected and incubated in permeabilization buffer (#554722; BD Biosciences) at 4°C for 20 min. After washing with Perm/Wash Buffer, cells were stained at 4°C for 30 min using the following primary antibodies: anti‐FOSL1 (#PA5‐76185; Thermo Fisher Scientific), CDK2 (#LS‐C351983; LSBio), CDK4 (#LS‐ C99873 ; LSBio), cyclin D (#LS‐B4507; LSBio), cyclin E (#32‐1600; Thermo Fisher Scientific), Oct4 (#ab184665; Abcam), Nanog (#ab109250; Abcam), Sox2 (#ab171380; Abcam), MGMT (#MA5‐13506; Thermo Fisher Scientific), IL‐6 (#M620; Thermo Fisher Scientific), STAT3 (#710077; Thermo Fisher Scientific), phospho‐STAT3 Ser727 (#PA5‐17876; Thermo Fisher Scientific), phospho‐STAT3 Tyr705 (#MA5‐15193; Thermo Fisher Scientific), and intracellular ROS (#ab113851; Abcam).

Techniques: Inhibition, Expressing, Derivative Assay, Flow Cytometry, Control, Quantitative Proteomics, Sublimation, Staining

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