p braf  (Cell Signaling Technology Inc)

Journal: International Journal of Molecular Medicine

Article Title: Novel Hsp90 inhibitor JD-02 inhibits HSV-1 infection via the Raf/MEK/ERK signaling pathway

doi: 10.3892/ijmm.2026.5810

Figure Lengend Snippet: JD-02 inhibits HSV-1 replication by suppressing the Raf/MEK/ERK signaling pathway. (A) Western blot analysis was conducted to assess the effects of HSV-1 (MOI=0.1) infection on the protein levels of BRAF, MEK and ERK, with and without treatment using JD-02. (B) Treatment with JD-02 at the specified concentration influences the protein levels of BRAF, MEK and ERK in HaCaT cells over a 12-h period. (C and D) HaCaT cells were subjected to transfection with either N.C. siRNA or ERK siRNA for a period of 48 h. Subsequently, the cells were infected with HSV-1 (MOI=0.1) for an additional 24 h. The DNA copy number of the viral gene UL54, as well as the viral protein expression of gB, ICP0, ICP27, ERK and p-ERK were evaluated. (E) The DNA copy numbers of the viral genes UL30 , UL52 and UL54 in HaCaT cells infected with HSV-1 (MOI=0.1) and subsequently treated with either JD-02 (1 μ M) or U0126 (10 μ M) for 24 h, were quantified using reverse transcription-quantitative PCR. (F) Western blot analysis of viral proteins (gB, ICP0 and ICP27), ERK and p-ERK expression in HaCaT cell infected with HSV-1 (MOI=0.1) and treated with U0126 for indicated concentration. (G) Western blot analysis of UL30 overexpression in HaCaT cells treated with U0126. Data are presented as the mean ± SD (n=3). * P<0.05, ** P<0.01, *** P<0.01 and **** P<0.0001 compared with the HSV-1 group. HSV, Herpes Simplex Virus; MOI, multiplicity of infection; N.C., negative control; siRNA, small interfering RNA; p-, phosphorylated.

Article Snippet: These primary antibodies included ICP0 (1:1,000; cat. no. ab6513; Abcam), ICP27 (1:1,000; cat. no. ab53480; Abcam), gD (1:1,000; cat. no. ab6507; Abcam), gB (1:500; cat. no. sc-56987; Santa Cruz Biotechnology, Inc.), Raf-B (1:500; cat. no. sc-166; Santa Cruz Biotechnology, Inc.), p-BRAF (1:1,000; cat. no. 2696T; Cell Signaling Technology, Inc.), ERK (1:1,000; cat. no. 4695S; Cell Signaling Technology, Inc.), p-ERK (1:1,000; cat. no. 4370S; Cell Signaling Technology, Inc.), MEK1/2 (1:1,000; cat. no. AF6385; Affinity Biosciences), p-MEK1/2 (1:1,000; cat. no. 9154T; Cell Signaling Technology, Inc.), Flag (1:1,000; cat. no. 14793S; Cell Signaling Technology, Inc.), HA (1:1,000; cat. no. 3724S; Cell Signaling Technology, Inc.).

Techniques: Western Blot, Infection, Concentration Assay, Transfection, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Over Expression, Virus, Negative Control, Small Interfering RNA

Journal: Cell Death & Disease

Article Title: OTUD4 promotes the progression of glioblastoma by deubiquitinating CDK1 and activating MAPK signaling pathway

doi: 10.1038/s41419-024-06569-x

Figure Lengend Snippet: Antibodies used in the experiment.

Article Snippet: p-BRAF(T401) , bs-5224R , , , BIOSS , Beijing, China.

Techniques: Western Blot

Journal: Biomedicines

Article Title: Activation of MAP Kinase Pathway by Polyisoprenylated Cysteinyl Amide Inhibitors Causes Apoptosis and Disrupts Breast Cancer Cell Invasion

doi: 10.3390/biomedicines12030470

Figure Lengend Snippet: Proposed mechanism of action of the PCAIs in breast cancer cell lines. Abbreviations: RTK, Receptor Tyrosine Kinase; GRB2, Growth Factor Receptor Bound Protein 2; SOS, Son-of-Sevenless; GDP, Guanosine Diphosphate; GTP, Guanosine Triphosphate; RAS, Rat Sarcoma; BRAF, Rapidly Accelerated Fibrosarcoma, v-Raf Murine Sarcoma Viral Oncogene Homolog B; CRAF, RAF Proto-Oncogene Serine/Threonine-Protein Kinase; MEK, Mitogen-Activated Protein Kinase Kinase; ERK, Extracellular-Signal-Regulated Kinases; p90RSK, 90 kDa Ribosomal s6 Kinases; AKT, Protein Kinase B; BAK1, BCL2 Antagonist/Killer 1.

Article Snippet: Primary antibodies specific to phosphorylated proteins BRAF (p-BRAF, Cat #2696), CRAF (p-CRAF, Cat #9427), MEK 1/2 (p-MEK 1/2, Cat #9154S), ERK 1/2 (p-ERK1/2, Cat #4370S), p90RSK (p-p90RSK, Cat #11989S), AKT (p-AKT, Cat # 4060s), vinculin (E1E9V, Cat #13901), full-length Caspase 3 (Cat #9662S), full-length Caspase 7 (Cat #9492S), secondary antibodies anti-mouse IgG, HRP-linked antibody (Cat #7076), and anti-rabbit IgG, HRP-linked antibody (Cat #7074), and housekeeping protein GAPDH (Cat #5174S) or α-actinin (Cat #6478S) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: