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Huabio Inc braf
Braf, supplied by Huabio Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/braf/anti+braf/pm42134490-238-44-45
Average 86 stars, based on 1 article reviews

braf - by Bioz Stars, 2026-10

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Article Title: ZDHHC5-mediated BRAF palmitoylation activates the MAPK pathway and drives cholangiocarcinoma progression.
Article Snippet: Cholangiocarcinoma is a highly malignant tumor with an increasing incidence around the world.. Discovery of novel molecular targets and effective therapies for cholangiocarcinoma are urgently needed.. Palmitoylation is a reversible lipid modification mainly catalyzed by ZDHHC family palmitoyltransferases.

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RKO and HT-29 cells exhibit intrinsic resistance to BRAFi monotherapy. (A) The effect of BRAFi (Encorafenib and Dabrafenib) on the growth of colon cancer cells, including Colo205, RKO, HT29 with BRAF V600E mutation, and HCT116, SW1463, LS513 with wild-type BRAF . The CRC cells were treated with BRAFi for 72 h, and the cell viability was determined by MTS assay. (B) The efficacy of BRAFi in suppressing the growth of BRAF V600E mutant CRC cells. Colo205, RKO, and HT-29 cells were seeded into 12 well plates and then exposed to BRAFi for 48 h. The living cells were stained by crystal violet. (C) The cytotoxic effect of BRAFi on clonogenic growth. 400 cells/well were seeded into the 6-well plates and then treated with BRAFi for 24 h. Fresh medium was exchanged every 3–4 days, and after 10–14 days the clones were stained by crystal violet. (D, E) the therapeutic efficacy of BRAFi on blocking p-ERK signal. Colo205, RKO, and HT-29 cells were treated with BRAFi alone with various dosing and timing, the p-ERK, ERK, and β -actin were determined by immunoblot analysis. A representative image from 3 to 5 independent experiments is shown.

Journal: Acta Pharmaceutica Sinica. B

Article Title: CDK9 inhibition sensitizes intrinsically resistant BRAF V600E mutant colorectal cancer to BRAF inhibitors

doi: 10.1016/j.apsb.2026.05.008

Figure Lengend Snippet: RKO and HT-29 cells exhibit intrinsic resistance to BRAFi monotherapy. (A) The effect of BRAFi (Encorafenib and Dabrafenib) on the growth of colon cancer cells, including Colo205, RKO, HT29 with BRAF V600E mutation, and HCT116, SW1463, LS513 with wild-type BRAF . The CRC cells were treated with BRAFi for 72 h, and the cell viability was determined by MTS assay. (B) The efficacy of BRAFi in suppressing the growth of BRAF V600E mutant CRC cells. Colo205, RKO, and HT-29 cells were seeded into 12 well plates and then exposed to BRAFi for 48 h. The living cells were stained by crystal violet. (C) The cytotoxic effect of BRAFi on clonogenic growth. 400 cells/well were seeded into the 6-well plates and then treated with BRAFi for 24 h. Fresh medium was exchanged every 3–4 days, and after 10–14 days the clones were stained by crystal violet. (D, E) the therapeutic efficacy of BRAFi on blocking p-ERK signal. Colo205, RKO, and HT-29 cells were treated with BRAFi alone with various dosing and timing, the p-ERK, ERK, and β -actin were determined by immunoblot analysis. A representative image from 3 to 5 independent experiments is shown.

Article Snippet: BRAF inhibitors (Encorafenib and Dabrafenib), 3 well-characterized CDK9 inhibitors (AZD4573, NVP2, BAY-1251152), MEK inhibitors (Binimetinib, Trametinib), ERK inhibitors (ASN007, Ravoxertinib), and the kinase library (Supporting Information ) were purchased from MedChemExpress LLC.

Techniques: Mutagenesis, MTS Assay, Staining, Clone Assay, Drug discovery, Blocking Assay, Western Blot

CDK9 was discovered as a candidate target by using high-throughput screening. (A) Kinase library screening was performed by HTS. (B) Network Cluster graph of the kinase library compared to the top hit compounds. Cluster nodes size based mean inhibition percentage in library screen. Each pathway (green) and target (pink) ( k = 0.7) compared to the top hit targets (blue) ( k = 0.5). (C) CDK9 inhibitor is the most potent candidate agent for enhancing the efficacy of BRAFi. (D, E) RNA-seq data showed that CDK9i suppresses the MAPK pathway, including BRAF , MAPK3 , MAP2K1 , MAPK1 , KRAS and EGFR . RKO cells were treated with NVP2 (30 nmol/L) for 24 h, and RNA-seq was performed. (F) The protein expression level of CDK9 and phosphorylation (ser2) of RNA polymerase II in human CRC cell lines (HCT116, RKO, HT-29, Colo205, LS513, LS174T and CCD841 cells) were determined by immunoblot analysis. (G) The perturbation effect of CDK9 inhibition on the proliferation of human CRC cells. (H) The effect of CDK9i on clonogenic growth. 400 cells/well were seeded into the 6-well plate, and then treated with CDK9i (AZD4573, NVP2 and BAY) for 24 h. After 10–14 days, clones (>50 cells) were fixed, stained by crystal violet, and manually counted. A representative of 3–5 experiments is shown. (I) Effect CDK9i on the growth of CRC cells. RKO, HT-29 and Colo205 cells were treated with CDK9i for 72 h, cell viability was determined by MTS assay, and the IC 50 was calculated by GraphPad Prism 9.0 software. Values represent the mean ± SD from 3 to 5 independent experiments.

Journal: Acta Pharmaceutica Sinica. B

Article Title: CDK9 inhibition sensitizes intrinsically resistant BRAF V600E mutant colorectal cancer to BRAF inhibitors

doi: 10.1016/j.apsb.2026.05.008

Figure Lengend Snippet: CDK9 was discovered as a candidate target by using high-throughput screening. (A) Kinase library screening was performed by HTS. (B) Network Cluster graph of the kinase library compared to the top hit compounds. Cluster nodes size based mean inhibition percentage in library screen. Each pathway (green) and target (pink) ( k = 0.7) compared to the top hit targets (blue) ( k = 0.5). (C) CDK9 inhibitor is the most potent candidate agent for enhancing the efficacy of BRAFi. (D, E) RNA-seq data showed that CDK9i suppresses the MAPK pathway, including BRAF , MAPK3 , MAP2K1 , MAPK1 , KRAS and EGFR . RKO cells were treated with NVP2 (30 nmol/L) for 24 h, and RNA-seq was performed. (F) The protein expression level of CDK9 and phosphorylation (ser2) of RNA polymerase II in human CRC cell lines (HCT116, RKO, HT-29, Colo205, LS513, LS174T and CCD841 cells) were determined by immunoblot analysis. (G) The perturbation effect of CDK9 inhibition on the proliferation of human CRC cells. (H) The effect of CDK9i on clonogenic growth. 400 cells/well were seeded into the 6-well plate, and then treated with CDK9i (AZD4573, NVP2 and BAY) for 24 h. After 10–14 days, clones (>50 cells) were fixed, stained by crystal violet, and manually counted. A representative of 3–5 experiments is shown. (I) Effect CDK9i on the growth of CRC cells. RKO, HT-29 and Colo205 cells were treated with CDK9i for 72 h, cell viability was determined by MTS assay, and the IC 50 was calculated by GraphPad Prism 9.0 software. Values represent the mean ± SD from 3 to 5 independent experiments.

Techniques: High Throughput Screening Assay, Library Screening, Inhibition, RNA Sequencing, Expressing, Phospho-proteomics, Western Blot, Clone Assay, Staining, MTS Assay, Software

The synergistic effect of CDK9i in combination with BRAFi, MEKi and ERKi in intrinsically resistant CRC cells. (A) The synergistic effect of CDK9i and BRAFi on the proliferation of BRAF V600E mutant human CRC cells. RKO and HT-29 cells were treated with CDK9 inhibitors (AZD4573, 10 nmol/L; NVP2, 30 nmol/L; BAY, 100 nmol/L) and/or BRAFi (Encorafenib; Dabrafenib, 300 nmol/L) for 48 h, then the living cells were stained by crystal violet. A representative of 3–5 experiments is shown. (B) Knock-in of BRAF V600E restores the synergistic effect of CDK9i and BRAFi on the growth of HCT116 cells. HCT116 cells with BRAF V600E mutation and parental cells were seeded in 12-well plate and treated with CDK9i and/or BRAFi for 48 h, and the living cells were stained by crystal violet, with a representative of 3–5 experiments shown. (C) The synergistic effect of CDK9i and BRAFi on the growth of BRAF V600E mutant human CRC cells. RKO or HT-29 cells were seeded in 96-well plates and then treated with different concentrations of CDK9i (NVP2, AZD4573, BAY) and/or BRAFi for 72 h. Cell growth was determined by the MTS assay. (D) Data from 3 to 5 independent experiments were used to calculate the Combination Index (CI) according to the Chou-Talalay method. (E, F) CDK9i enhances the efficacy of BRAFi on inducing apoptosis. RKO cells were exposed to CDK9i and/or BRAFi for 24 h, the apoptotic cells were measured by flow cytometry. (G) RKO cells were treated with CDK9i and/or BRAFi for 24 h, the cleaved PARP were determined by immunoblot analysis. (H) CDK9 knockdown enhance the effect of BRAFi on inducing cleaved PARP. (I) BRAF V600E knock-in restore the synergistic effect of CDK9i and BRAFi on inducing programmed cell death. (J) Non-cytotoxic effect of CDK9i and/or BRAFi on the growth of normal human colon epithelial cells. (K, L) CDK9i enhanced the efficacy of MEKi and ERKi. RKO cells were treated with CDK9i and/or MEKi (Binimetinib or Trametinib)/ERKi (ASN007 or Ravo) for 72 h, the cell growth was determined by MTS assay. The cells were treated with CDK9i and/or MEKi/ERKi for 24 h, then cleaved PARP and β -actin were determined by immunoblot analysis. Values in the graph represent the mean ± SD from 3 to 5 independent experiments. ∗∗ P < 0.01, vs. control.

Journal: Acta Pharmaceutica Sinica. B

Article Title: CDK9 inhibition sensitizes intrinsically resistant BRAF V600E mutant colorectal cancer to BRAF inhibitors

doi: 10.1016/j.apsb.2026.05.008

Figure Lengend Snippet: The synergistic effect of CDK9i in combination with BRAFi, MEKi and ERKi in intrinsically resistant CRC cells. (A) The synergistic effect of CDK9i and BRAFi on the proliferation of BRAF V600E mutant human CRC cells. RKO and HT-29 cells were treated with CDK9 inhibitors (AZD4573, 10 nmol/L; NVP2, 30 nmol/L; BAY, 100 nmol/L) and/or BRAFi (Encorafenib; Dabrafenib, 300 nmol/L) for 48 h, then the living cells were stained by crystal violet. A representative of 3–5 experiments is shown. (B) Knock-in of BRAF V600E restores the synergistic effect of CDK9i and BRAFi on the growth of HCT116 cells. HCT116 cells with BRAF V600E mutation and parental cells were seeded in 12-well plate and treated with CDK9i and/or BRAFi for 48 h, and the living cells were stained by crystal violet, with a representative of 3–5 experiments shown. (C) The synergistic effect of CDK9i and BRAFi on the growth of BRAF V600E mutant human CRC cells. RKO or HT-29 cells were seeded in 96-well plates and then treated with different concentrations of CDK9i (NVP2, AZD4573, BAY) and/or BRAFi for 72 h. Cell growth was determined by the MTS assay. (D) Data from 3 to 5 independent experiments were used to calculate the Combination Index (CI) according to the Chou-Talalay method. (E, F) CDK9i enhances the efficacy of BRAFi on inducing apoptosis. RKO cells were exposed to CDK9i and/or BRAFi for 24 h, the apoptotic cells were measured by flow cytometry. (G) RKO cells were treated with CDK9i and/or BRAFi for 24 h, the cleaved PARP were determined by immunoblot analysis. (H) CDK9 knockdown enhance the effect of BRAFi on inducing cleaved PARP. (I) BRAF V600E knock-in restore the synergistic effect of CDK9i and BRAFi on inducing programmed cell death. (J) Non-cytotoxic effect of CDK9i and/or BRAFi on the growth of normal human colon epithelial cells. (K, L) CDK9i enhanced the efficacy of MEKi and ERKi. RKO cells were treated with CDK9i and/or MEKi (Binimetinib or Trametinib)/ERKi (ASN007 or Ravo) for 72 h, the cell growth was determined by MTS assay. The cells were treated with CDK9i and/or MEKi/ERKi for 24 h, then cleaved PARP and β -actin were determined by immunoblot analysis. Values in the graph represent the mean ± SD from 3 to 5 independent experiments. ∗∗ P < 0.01, vs. control.

Techniques: Mutagenesis, Staining, Knock-In, MTS Assay, Flow Cytometry, Western Blot, Knockdown, Control

CDK9i synergistically blocked multiple cancer signaling pathways and enhanced the therapeutic response of BRAFi. (A) The down-regulated target proteins in the cancer signaling network after treatment with CDK9i and BRAFi. RKO cells were treated with NVP2 (30 nmol/L) and/or Encorafenib (300 nmol/L) for 24 h, then cell lysis was performed for RPPA analysis. (B) The up-regulated target proteins in cancer signaling network after exposure to CDK9i in combination with BRAFi. (C) The effect of this combination on the alteration biological processes in human cancer. (D) The synergistic effect of CDK9i and BRAFi on inhibition of BRAF/MEK/ERK/S6 pathway. (E–G) CDK9i enhances the efficacy of BRAFi on blocking p-ERK/p-S6 and inducing cleaved PARP. (H) The synergistic effect of CDK9i and BRAFi on suppression of AKT/mTOR/4 EB-P1 pathway. (I) CDK9i enhances the effect of BRAFi on mitochondria-mediated cellular apoptosis in intrinsic resistance CRC cells. (J) Signaling pathways altered by the combination of CDK9i and BRAFi (Created in BioRender. Wei, N (2026) https://BioRender.com/m2l5b57 ). The combination of CDK9i and BRAFi leads to synergistic inhibition of oncogenic signaling pathways, including BRAF/MEK/ERK/S6, AKT/mTOR/4E-BP1, and induction of mitochondria-mediated apoptosis. All protein expression levels of targets were determined by immunoblot analysis. A representative image from 3 to 5 independent experiments is shown. Values in the graph represent the mean ± SD from 3 to 5 independent experiments. ∗∗∗ P < 0.001 vs. control.

Journal: Acta Pharmaceutica Sinica. B

Article Title: CDK9 inhibition sensitizes intrinsically resistant BRAF V600E mutant colorectal cancer to BRAF inhibitors

doi: 10.1016/j.apsb.2026.05.008

Figure Lengend Snippet: CDK9i synergistically blocked multiple cancer signaling pathways and enhanced the therapeutic response of BRAFi. (A) The down-regulated target proteins in the cancer signaling network after treatment with CDK9i and BRAFi. RKO cells were treated with NVP2 (30 nmol/L) and/or Encorafenib (300 nmol/L) for 24 h, then cell lysis was performed for RPPA analysis. (B) The up-regulated target proteins in cancer signaling network after exposure to CDK9i in combination with BRAFi. (C) The effect of this combination on the alteration biological processes in human cancer. (D) The synergistic effect of CDK9i and BRAFi on inhibition of BRAF/MEK/ERK/S6 pathway. (E–G) CDK9i enhances the efficacy of BRAFi on blocking p-ERK/p-S6 and inducing cleaved PARP. (H) The synergistic effect of CDK9i and BRAFi on suppression of AKT/mTOR/4 EB-P1 pathway. (I) CDK9i enhances the effect of BRAFi on mitochondria-mediated cellular apoptosis in intrinsic resistance CRC cells. (J) Signaling pathways altered by the combination of CDK9i and BRAFi (Created in BioRender. Wei, N (2026) https://BioRender.com/m2l5b57 ). The combination of CDK9i and BRAFi leads to synergistic inhibition of oncogenic signaling pathways, including BRAF/MEK/ERK/S6, AKT/mTOR/4E-BP1, and induction of mitochondria-mediated apoptosis. All protein expression levels of targets were determined by immunoblot analysis. A representative image from 3 to 5 independent experiments is shown. Values in the graph represent the mean ± SD from 3 to 5 independent experiments. ∗∗∗ P < 0.001 vs. control.

Techniques: Protein-Protein interactions, Clinical Proteomics, Lysis, Inhibition, Blocking Assay, Expressing, Western Blot, Control

Synergistic effect of CDK9i and BRAFi on the growth of CRC PDOs with BRAF V600E mutation, but not BRAF wild type PDOs. (A) IHC staining of PDO CK9677, including Ki-67, cleave caspase-3, and p-ERK. Scale bar: 100 μm. (B) The synergistic effect of CDK9i and/or BRAFi on the growth of CRC PDO CK9677. CK9677 ( BRAF V600E ) PDO were treated with NVP2 and/or Encorafenib for 7 days. PDO viability was determined using CellTiter Glo 3D assay; (C) The combination index was calculated by using the Chou-Talalay method. (D) The synergistic effect of CDK9i and BRAFi on suppressing p-ERK/p-S6 and C-Myc, inducing cleaved PARP in PDO. (E) Therapeutic response to the CDK9i in combination with BRAFi in multiple PDOs, including BRAF V600E (CK7719, CK8485, CK8911); BRAF WT (CRC-22-008 and CK10278). The organoids were exposed to CDK9i and/BRAFi for 5 days, and images were shown at Day 0 and Day 5; After 7 days treatment, the growth of PDOs were determined using CellTiter Glo 3D assay. Values in the graph represent the mean ± SD from 3 to 5 independent experiments. ∗ P<0.05 , ∗∗ P < 0.01; N.S. (not significant) NVP2 vs. Combination; # means CI > 2.

Journal: Acta Pharmaceutica Sinica. B

Article Title: CDK9 inhibition sensitizes intrinsically resistant BRAF V600E mutant colorectal cancer to BRAF inhibitors

doi: 10.1016/j.apsb.2026.05.008

Figure Lengend Snippet: Synergistic effect of CDK9i and BRAFi on the growth of CRC PDOs with BRAF V600E mutation, but not BRAF wild type PDOs. (A) IHC staining of PDO CK9677, including Ki-67, cleave caspase-3, and p-ERK. Scale bar: 100 μm. (B) The synergistic effect of CDK9i and/or BRAFi on the growth of CRC PDO CK9677. CK9677 ( BRAF V600E ) PDO were treated with NVP2 and/or Encorafenib for 7 days. PDO viability was determined using CellTiter Glo 3D assay; (C) The combination index was calculated by using the Chou-Talalay method. (D) The synergistic effect of CDK9i and BRAFi on suppressing p-ERK/p-S6 and C-Myc, inducing cleaved PARP in PDO. (E) Therapeutic response to the CDK9i in combination with BRAFi in multiple PDOs, including BRAF V600E (CK7719, CK8485, CK8911); BRAF WT (CRC-22-008 and CK10278). The organoids were exposed to CDK9i and/BRAFi for 5 days, and images were shown at Day 0 and Day 5; After 7 days treatment, the growth of PDOs were determined using CellTiter Glo 3D assay. Values in the graph represent the mean ± SD from 3 to 5 independent experiments. ∗ P<0.05 , ∗∗ P < 0.01; N.S. (not significant) NVP2 vs. Combination; # means CI > 2.

Techniques: Mutagenesis, Immunohistochemistry, Clinical Proteomics

CDK9i sensitizes the in vivo therapeutic efficacy of BRAFi in RKO-xenograft and PDX with BRAF V600E . The synergistic effect of Encorafenib and NVP2 on tumor growth. Athymic nude mice bearing RKO tumor xenografts were administered 5 daily/week vehicle control, Encorafenib (10 mg/kg), NVP2 (8 mg/kg), and combination orally for 3 weeks (5 mice per group). (A, D) Tumor volume and body weight were measured 3 times per week. (B) The image of excised tumor and tumor weights (C) after treatment with CDK9i and/or BRAFi. (E, F) Tumors were fixed in formalin and paraffin-embedded. Slides were stained for Ki-67, TUNEL and p-ERK; NOD-SCID mice bearing BRAF V600E mutant PDX model. After tumor size reached ∼300 mm 3 , the mice were treated with vehicle control, NPV2 and/or Encoranfenib orally for 5 days/week for 20 days (5 mice/group). (G) Tumor volume and body weight (H) were monitored 3 times per week; (I, J) protein expression in BRAF V600E PDX was determined after 20 days of CDK9i and/or BRAFi treatment. Tumors were harvested 2 h after the last dose. (K, L) Tumors were fixed in formalin and paraffin-embedded. Slides were stained for Ki-67, TUNEL and p-S6. Scale bar: 50 μm; After tumor size reached ∼200 mm 3 , the mice bearing BRAF WT CRC tumor (CM0050) were treated with vehicle control, NPV2 and/or Encorafenib orally 5 days/week for 30 days. (M) Tumor size and (N) body weight of PDX mice were monitored; Values in the graph represent the mean ± SD. ∗∗∗ P < 0.001 CDK9i (NVP2) vs. Combination (NVP2+Enco); N.S. P > 0.05 CDK9i (NVP2) vs. Combination (NVP2+Enco).

Journal: Acta Pharmaceutica Sinica. B

Article Title: CDK9 inhibition sensitizes intrinsically resistant BRAF V600E mutant colorectal cancer to BRAF inhibitors

doi: 10.1016/j.apsb.2026.05.008

Figure Lengend Snippet: CDK9i sensitizes the in vivo therapeutic efficacy of BRAFi in RKO-xenograft and PDX with BRAF V600E . The synergistic effect of Encorafenib and NVP2 on tumor growth. Athymic nude mice bearing RKO tumor xenografts were administered 5 daily/week vehicle control, Encorafenib (10 mg/kg), NVP2 (8 mg/kg), and combination orally for 3 weeks (5 mice per group). (A, D) Tumor volume and body weight were measured 3 times per week. (B) The image of excised tumor and tumor weights (C) after treatment with CDK9i and/or BRAFi. (E, F) Tumors were fixed in formalin and paraffin-embedded. Slides were stained for Ki-67, TUNEL and p-ERK; NOD-SCID mice bearing BRAF V600E mutant PDX model. After tumor size reached ∼300 mm 3 , the mice were treated with vehicle control, NPV2 and/or Encoranfenib orally for 5 days/week for 20 days (5 mice/group). (G) Tumor volume and body weight (H) were monitored 3 times per week; (I, J) protein expression in BRAF V600E PDX was determined after 20 days of CDK9i and/or BRAFi treatment. Tumors were harvested 2 h after the last dose. (K, L) Tumors were fixed in formalin and paraffin-embedded. Slides were stained for Ki-67, TUNEL and p-S6. Scale bar: 50 μm; After tumor size reached ∼200 mm 3 , the mice bearing BRAF WT CRC tumor (CM0050) were treated with vehicle control, NPV2 and/or Encorafenib orally 5 days/week for 30 days. (M) Tumor size and (N) body weight of PDX mice were monitored; Values in the graph represent the mean ± SD. ∗∗∗ P < 0.001 CDK9i (NVP2) vs. Combination (NVP2+Enco); N.S. P > 0.05 CDK9i (NVP2) vs. Combination (NVP2+Enco).

Techniques: In Vivo, Drug discovery, Control, Staining, TUNEL Assay, Mutagenesis, Expressing

CDK9i in combination with BRAFi exerts synergistic effect in chemoresistant CRC. (A) The effect of 5-FU on the growth of RKO-5FUR10 and parental cells. RKO and RKO-5FUR10 cells were treated with 5-FU for 72 h, and the cell viability was determined by MTS assay; (B) The cytotoxic effect 5-FU on the survival of RKO and RKO-5FUR10 cells; RKO or RKO-5FUR10 cells were incubated with 5-FU for 48 h, the living cells were stained by crystal violet. (C) The efficacy of FOLFOX in combination with BRAFi on the growth of RKO and RKO-5FUR10 cells; (D, E) The synergistic effect of CDK9i and BRAFi on the growth of RKO-5FUR10 cells. RKO-5FUR10 cells were treated with CDK9i (NVP2) and/or BRAFi for 48 or 72 h. Cell growth was visualized and quantified by crystal violet and the MTS assay, respectively. (F) Data from 3 to 5 independent experiments were used to calculate the Combination Index (CI). (G, H) The effect of CDK9i and BRAFi on alteration of cancer signaling network, including BRAF/MEK/ERK/S6; AKT/mTOR/4E-BP1 and downstream effectors of CDK9. RKO-5FUR10 cells were treated with CDK9i and/or BRAFi for 24 h, the expression level of relevant target proteins was determined by immunoblot analysis. (I) The cytotoxic effect of 5-FU on the growth of PDOs with BRAF V600E . After treatment with 5-FU for 7 days, the viability of PDOs was determined by Cell Titer Glo 3D. (J) The synergistic effect of CDK9i and BRAFi on suppressing p-ERK in 5-FU-resistant PDO (CK7719); A representative image was shown from 3 independent experiments.

Journal: Acta Pharmaceutica Sinica. B

Article Title: CDK9 inhibition sensitizes intrinsically resistant BRAF V600E mutant colorectal cancer to BRAF inhibitors

doi: 10.1016/j.apsb.2026.05.008

Figure Lengend Snippet: CDK9i in combination with BRAFi exerts synergistic effect in chemoresistant CRC. (A) The effect of 5-FU on the growth of RKO-5FUR10 and parental cells. RKO and RKO-5FUR10 cells were treated with 5-FU for 72 h, and the cell viability was determined by MTS assay; (B) The cytotoxic effect 5-FU on the survival of RKO and RKO-5FUR10 cells; RKO or RKO-5FUR10 cells were incubated with 5-FU for 48 h, the living cells were stained by crystal violet. (C) The efficacy of FOLFOX in combination with BRAFi on the growth of RKO and RKO-5FUR10 cells; (D, E) The synergistic effect of CDK9i and BRAFi on the growth of RKO-5FUR10 cells. RKO-5FUR10 cells were treated with CDK9i (NVP2) and/or BRAFi for 48 or 72 h. Cell growth was visualized and quantified by crystal violet and the MTS assay, respectively. (F) Data from 3 to 5 independent experiments were used to calculate the Combination Index (CI). (G, H) The effect of CDK9i and BRAFi on alteration of cancer signaling network, including BRAF/MEK/ERK/S6; AKT/mTOR/4E-BP1 and downstream effectors of CDK9. RKO-5FUR10 cells were treated with CDK9i and/or BRAFi for 24 h, the expression level of relevant target proteins was determined by immunoblot analysis. (I) The cytotoxic effect of 5-FU on the growth of PDOs with BRAF V600E . After treatment with 5-FU for 7 days, the viability of PDOs was determined by Cell Titer Glo 3D. (J) The synergistic effect of CDK9i and BRAFi on suppressing p-ERK in 5-FU-resistant PDO (CK7719); A representative image was shown from 3 independent experiments.

Techniques: MTS Assay, Incubation, Staining, Expressing, Western Blot

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.

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

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.

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.

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

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.

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.

Techniques: Staining, Negative Control, Over Expression

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.

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

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.

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

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