Journal: Molecular Medicine Reports

Article Title: RUVBL1 and CRAF promote periodontal ligament stem cell osteogenic differentiation via the MEK/ERK signaling cascade

doi: 10.3892/mmr.2026.13881

Figure Lengend Snippet: Overexpression of CRAF activates the RAF/MEK/ERK signaling pathway. Reverse transcription-quantitative PCR was used to analyze the mRNA expression of (A) ARAF, (B) BRAF, (C) CRAF, (D) MEK1, (E) MEK2, (F) ERK1and (G) ERK2. Western blotting was used to analyze the protein levels of (H) ARAF, (I) BRAF, (J) CRAF, (K) MEK1/2, (L) p-MEK1/2, (M) p-MEK1/2 to MEK1/2, (N) ERK1/2, (O) p-ERK1/2 and (P) p-ERK1/2 to ERK1/2. *P<0.05, **P<0.01 and ***P<0.001. p-, phosphorylated; OE, overexpression; NC, negative control; sh, short hairpin; ns, not significant; ARAF, A-Raf proto-oncogene serine/threonine-protein kinase; BRAF, B-Raf proto-oncogene serine/threonine-protein kinase; CRAF, C-Raf proto-oncogene serine/threonine-protein kinase.

Article Snippet: The antibodies were as follows: GAPDH (1:1,000; cat. no. P30008M; Abmart Pharmaceutical Technology Co., Ltd.), A-Raf proto-oncogene serine/threonine-protein kinase (ARAF) (cat. no. bs-2251R), CRAF (cat. no. bs-23170R), MEK1/2 (cat. no. bs-1041R), phosphorylated (p)-MEK1/2 (cat. no. bs-3270R), ERK1/2 (cat. no. bsm-33232M), p-ERK1/2 (cat. no. bs-3016R; all BIOSS), RUVBL1 (cat. no. 74775; Cell Signaling Technology, Inc.), ERK1/2 (all 1:500; cat. no. bsm-33232M; BIOSS), B-Raf proto-oncogene serine/threonine-protein kinase (BRAF) (1:2,000; cat. no. ab33899; Abcam) and HRP-conjugated universal secondary antibody [cat. nos.

Techniques: Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Negative Control

Journal: Molecular Medicine Reports

Article Title: RUVBL1 and CRAF promote periodontal ligament stem cell osteogenic differentiation via the MEK/ERK signaling cascade

doi: 10.3892/mmr.2026.13881

Figure Lengend Snippet: Overexpression of RUVBL1 activates the MEK/ERK signaling pathway. Reverse transcription-quantitative PCR was used to analyze the mRNA expression of (A) ARAF, (B) BRAF, (C) CRAF, (D) MEK1, (E) MEK2, (F) ERK1 and (G) ERK2. Western blotting was used to analyze protein levels of (H) ARAF, (I) BRAF, (J) CRAF, (K) MEK1/2, (L) p-MEK1/2, (M) p-MEK1/2 to MEK1/2, (N) ERK1/2, (O) p-ERK1/2 and (P) p-ERK1/2 to ERK1/2. *P<0.05, **P<0.01 and ***P<0.001. RUVBL1, RuvB-like AAA ATPase-1; p-, phosphorylated; OE, overexpression; NC, negative control; sh, short hairpin; ns, not significant; ARAF, A-Raf proto-oncogene serine/threonine-protein kinase; BRAF, B-Raf proto-oncogene serine/threonine-protein kinase; CRAF, C-Raf proto-oncogene serine/threonine-protein kinase.

Article Snippet: The antibodies were as follows: GAPDH (1:1,000; cat. no. P30008M; Abmart Pharmaceutical Technology Co., Ltd.), A-Raf proto-oncogene serine/threonine-protein kinase (ARAF) (cat. no. bs-2251R), CRAF (cat. no. bs-23170R), MEK1/2 (cat. no. bs-1041R), phosphorylated (p)-MEK1/2 (cat. no. bs-3270R), ERK1/2 (cat. no. bsm-33232M), p-ERK1/2 (cat. no. bs-3016R; all BIOSS), RUVBL1 (cat. no. 74775; Cell Signaling Technology, Inc.), ERK1/2 (all 1:500; cat. no. bsm-33232M; BIOSS), B-Raf proto-oncogene serine/threonine-protein kinase (BRAF) (1:2,000; cat. no. ab33899; Abcam) and HRP-conjugated universal secondary antibody [cat. nos.

Techniques: Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Negative Control

Journal: Molecular Medicine Reports

Article Title: RUVBL1 and CRAF promote periodontal ligament stem cell osteogenic differentiation via the MEK/ERK signaling cascade

doi: 10.3892/mmr.2026.13881

Figure Lengend Snippet: ODN MT01 promotes osteogenic differentiation of PDLSCs. (A) PDLSC proliferation and (B) ALP staining following treatment with varying concentrations of ODN MT01. (C) Osteogenic differentiation and (D) mineralization after the addition of ODN MT01 and osteogenic inducers. ALP staining was used to assess the effect of (E) CRAF and (F) RUVBL1 on osteogenic differentiation of PDLSCs following the addition of ODN MT01 and osteogenic inducers. Alizarin Red staining was used to assess the effect of (G) CRAF and (H) RUVBL1 on the degree of mineralization of PDLSCs following the addition of ODN MT01 and osteogenic inducers. *P<0.05 and ***P<0.001. ODN, oligodeoxynucleotide; PDLSC, periodontal ligament stem cell; ALP, alkaline phosphatase, RUVBL1, RuvB-like AAA ATPase-1; NC, negative control; OE, overexpression; sh, short hairpin; ns, not significant; CRAF, C-Raf proto-oncogene serine/threonine-protein kinase; OI, osteogenic induction; OM, ODN MT01.

Article Snippet: The antibodies were as follows: GAPDH (1:1,000; cat. no. P30008M; Abmart Pharmaceutical Technology Co., Ltd.), A-Raf proto-oncogene serine/threonine-protein kinase (ARAF) (cat. no. bs-2251R), CRAF (cat. no. bs-23170R), MEK1/2 (cat. no. bs-1041R), phosphorylated (p)-MEK1/2 (cat. no. bs-3270R), ERK1/2 (cat. no. bsm-33232M), p-ERK1/2 (cat. no. bs-3016R; all BIOSS), RUVBL1 (cat. no. 74775; Cell Signaling Technology, Inc.), ERK1/2 (all 1:500; cat. no. bsm-33232M; BIOSS), B-Raf proto-oncogene serine/threonine-protein kinase (BRAF) (1:2,000; cat. no. ab33899; Abcam) and HRP-conjugated universal secondary antibody [cat. nos.

Techniques: Staining, Negative Control, Over Expression

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: iScience

Article Title: RIPOR2 promotes multinucleation of melanoma cells downstream of the RAS/ERK oncogenic pathway

doi: 10.1016/j.isci.2026.115734

Figure Lengend Snippet: RIPOR2 is expressed in the melanocytes of an intermediate-grade melanocytic lesion Adjacent sections of an early BRAF V600E -positive intraepidermal melanoma. Dotted boxes are magnified in the adjacent images. (A) H&E stain shows tissue disorganization in the center of the section. (B) Immunohistochemistry with an anti-BRAF V600E antibody stains the mutated melanocytes and highlights the malignant lesion zone. (C) Immunofluorescence with antiSOX10. (D) Anti-RIPOR2 antibodies demonstrated that RIPOR2 is expressed in SOX10+ epidermal nests of BRAF V600E -positive melanocytes, suggesting that it is expressed in malignant melanocytes. The RIPOR2 staining is cytoplasmic. Blue corresponds to Hoechst nuclear staining in all panels. Scale bars, 100 μm.

Article Snippet: 3.5 μm thick paraffine sections were done from samples and staining was done on a Benchmark ULTRA immunohistochemistry automated system (Ventana, Roche Diagnostics), using the anti-BRAF antibody directed against the V600E mutant form (clone VE1, Eurobio Scientific), dilution 1:250, Optiview DAB detection (Ventana).

Techniques: Staining, Immunohistochemistry, Immunofluorescence