u 87 mg luc2  (ATCC)

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: Firefly luciferase‐expressing U87MG cells (U87MG/Luc2; ATCC #HTB‐14‐LUC2) were also utilized and cultured in RPMI‐1640 medium supplemented with 10% FBS, 1% penicillin‐streptomycin, and 8 μg/mL blasticidin (all from Thermo Fisher Scientific).

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

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 reduces the stemness hallmark. (A) Representative histogram of protein expression, including MGMT and stemness hallmarks, in GBM patient's cells was shown (top panel). Protein expression in patients with GBM‐derived cells ( n = 6), including MGMT and stemness hallmarks, analyzed using flow cytometry and quantified using FlowJo V10 (bottom panel). (B) Expression of stemness‐related proteins in U87MG cells analyzed using flow cytometry and quantified using FlowJo V10. (C) Representative images of the wound healing analysis of U87MG cells treated with si‐control, si‐FOSL1, or TMZ (0, 6, and 24 h). Scale bars, 200 µm. (D) Quantification of the cell wound area at each time point in U87MG cells was conducted with ImageJ. *p < 0.05; **p < 0.005; ***p < 0.0005; paired t ‐test ( n = 3).

Article Snippet: Firefly luciferase‐expressing U87MG cells (U87MG/Luc2; ATCC #HTB‐14‐LUC2) were also utilized and cultured in RPMI‐1640 medium supplemented with 10% FBS, 1% penicillin‐streptomycin, and 8 μg/mL blasticidin (all from Thermo Fisher Scientific).

Techniques: Inhibition, Expressing, Derivative Assay, Flow Cytometry, Control

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: FOSL1 regulates the IL‐6/STAT3 signaling pathway, leading to GBM stemness. (A) Representative histogram for flow cytometry analysis of the IL‐6/STAT3 axis in U87MG cells (gray: unstained, red: si‐control, blue: si‐FOSL1, top panel). δ‐MFI values were shown on the histogram. Quantification of IL‐6/STAT3 signaling pathway‐associated molecules measured using flow cytometry in U87MG cells (bottom panel). (B) Quantification of IL‐6/STAT3 signaling pathway‐associated molecules measured using flow cytometry in U87MG cells. The data are shown as δ‐MFI values. (C) Quantification of stemness hallmarks measured using flow cytometry in U87MG cells. *p < 0.05; **p < 0.005; ***p < 0.0005; paired t ‐test ( n = 3).

Article Snippet: Firefly luciferase‐expressing U87MG cells (U87MG/Luc2; ATCC #HTB‐14‐LUC2) were also utilized and cultured in RPMI‐1640 medium supplemented with 10% FBS, 1% penicillin‐streptomycin, and 8 μg/mL blasticidin (all from Thermo Fisher Scientific).

Techniques: Flow Cytometry, Control

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: Vemurafenib sensitizes TMZ responsiveness via FOSL1 downregulation in GBM cells. (A, B) Each protein, including FOSL1 and stemness hallmark, was analyzed using flow cytometry, and δ‐MFI of proteins was quantified using FlowJo V10. C Viability of U87MG treated with TMZ or VEM was quantified by WST‐8 reduction analysis following the manufacturer's instruction. (D) Image of tumor specimens from xenograft BALB/c nude mice model. Scale bar, 10 mm. (E) Measurement of tumor volume and relative tumor volume in xenograft GBM mice for 21 days. Tumor volume (mm 3 ) was measured with calipers at the indicated time points. Tumor volume (mm 3 ) = d 2 × D /2, where d and D are the shortest and longest diameters (mm). (F) Measurement of mouse and tumor weight. (G) Kaplan–Meier curve for the orthotopic GBM mouse model treated with TMZ or VEM. (H) Representative image of immunofluorescence for FOSL1 and TUNEL expression in GBM tissues of orthotopic nude mice (blue: DAPI, red: FOSL1, green: TUNEL). Scale bars, 50 µm. (I) Expression of FOSL1 in orthotopic GBM mouse tissue quantified using ImageJ. (J) Quantification of TUNEL staining in orthotopic GBM mouse tissues. The stained cells were quantified using ImageJ. *p < 0.05; **p < 0.005; ***p < 0.0005; paired t ‐test (xenograft model, n = 36; orthotopic model, n = 28).

Article Snippet: Firefly luciferase‐expressing U87MG cells (U87MG/Luc2; ATCC #HTB‐14‐LUC2) were also utilized and cultured in RPMI‐1640 medium supplemented with 10% FBS, 1% penicillin‐streptomycin, and 8 μg/mL blasticidin (all from Thermo Fisher Scientific).

Techniques: Flow Cytometry, Immunofluorescence, TUNEL Assay, Expressing, Staining