Journal: Poultry Science

Article Title: Curcumin alleviates proliferation dysfunction in chicken granulosa cells under oxidative stress through AKT-Raf1-ERK1/2 signaling pathway

doi: 10.1016/j.psj.2025.105342

Figure Lengend Snippet: Curcumin modulates the AKT-Raf1-ERK1/2 signaling pathway in chicken granulosa cells under oxidative stress. (A) Protein expression of AKT1, Raf1, and ERK1/2 under oxidative stress and curcumin treatment. (B) Relative expression of AKT. (C) Relative expression of p-AKT. (D-E) Changes in the relative expression and phosphorylation levels of Raf1. (F-G) Changes in the relative expression and phosphorylation levels of ERK1/2. The results are presented in the form of mean ± SD. n = 4. * p < 0.05; ** p < 0.01; **** p < 0.0001; ns p ≥ 0.05.

Article Snippet: ZenBio , Raf1 Rabbit pAb , 1: 1000.

Techniques: Expressing, Phospho-proteomics

Journal: Cell Death Discovery

Article Title: Dual role of PID1 in regulating apoptosis induced by distinct anticancer-agents through AKT/Raf-1-dependent pathway in hepatocellular carcinoma

doi: 10.1038/s41420-023-01405-1

Figure Lengend Snippet: A Cell apoptosis in HepG2 cells and Hepa1–6 cells with or without PID1 overexpression upon H 2 O 2 (200 μM) treatment for 24 h in the presence of Venetoclax (Ven) (10 μM). B BAD location, cytochrome c release and caspase-3 activation in response to H 2 O 2 (200 μM) in HepG2 cells and Hepa1–6 cells with or without PID1 overexpression. Cytoplasmic fraction and mitochondrial fraction in cells were prepared. C Immunofluorescence staining of Tom20 and BAD in Hep3B cells upon indicated treatments. Merged images show the overlap of Tom20 (red), BAD (green) and nuclear staining by DAPI (blue). D Immunofluorescence staining of Tom20 and cytochrome c in Hep3B cells upon indicated treatments. Merged images show the overlap of Tom20 (red), cytochrome c (green) and nuclear staining by DAPI (blue). E Western blot analysis of Raf-1, p-Raf-1 S338 , p-Raf-1 S259 , AKT, p-AKT T308 and β-actin in HepG2 cells and Hepa1–6 cells with or without PID1 overexpression. F Western blot analysis of Raf-1, p-Raf-1 S338 , p-Raf-1 S259 , AKT, p-AKT T308 and β-actin in HepG2 cells and Hepa1–6 cells with or without PID1 overexpression in the presence of AKT inhibitor VIII (iAKT) (10 μM). G Cell apoptosis in HepG2 cells and Hepa1–6 cells with or without PID1 overexpression upon Doxorubicin (200 nM) treatment for 24 h in the presence of AKT inhibitor VIII (10 μM). H Mitochondrial membrane potential in HepG2 cells and Hepa1–6 cells with or without PID1 overexpression upon Doxorubicin (200 nM) treatment for 24 h in the presence of AKT inhibitor VIII (10 μM) was examined by cytometry analysis, and Red/Green ratio was calculated. I Intracellular ROS in HepG2 cells and Hepa1–6 cells with or without PID1 overexpression upon Doxorubicin (200 nM) treatment for 24 h in the presence of AKT inhibitor VIII (10 μM) was examined by cytometry analysis, and MFI was calculated. J HepG2 cells with or without PID1 overexpression were treated with AKT inhibitor VIII (10 μM) and mitochondria-located BAD levels were detected by western blot analysis. Cytoplasmic fraction and mitochondrial fraction in cells were prepared. K In vitro kinase assay was performed with bacterially purified His-BAD as substrate, and immunoprecipitated HA-Raf-1 in HepG2 cells with or without PID1 overexpression upon AKT inhibitor VIII treatment as kinase. Phosphorylation of His-BAD at Ser112 was detected by western blot analysis. L Cell apoptosis in Hep3B cells with or without PID1 knockout combined with Raf-1 knockdown upon Doxorubicin (200 nM) treatment for 24 h was examined by cytometry analysis. Data are expressed as mean ± SD ( n = 3). * p < 0.05; ** p < 0.01; *** p < 0.001, ns not significant.

Article Snippet: Antibodies used to determine the protein expression were the following: PID1 (#27951; Signalway Antibody, Greenbelt, Maryland, USA), β-actin (#66009-1-lg; Proteintech, USA), Bcl-xL (#2764; Cell Signaling Technology, USA), Bcl2 (#12789-1-AP; Proteintech), BAD (#sc-8044 Santa Cruz Technology, Texas, USA), p-BAD S112 (#5284; Cell Signaling Technology), p-BAD S136 (#4366; Cell Signaling Technology), cytochrome c (#sc-13156; Santa Cruz Technology), cleaved caspase-3 (#9664; Cell Signaling Technology), Tom20 (#42406; Cell Signaling Technology), Raf-1 (#9422; Cell Signaling Technology), p-Raf-1 S259 (#9421; Cell Signaling Technology), p-Raf-1 S338 (#9427; Cell Signaling Technology), AKT (#4691; Cell Signaling Technology), p-AKT T308 (#13038; Cell Signaling Technology), HA-tag (#sc-7392; Santa Cruz Technology), His-tag (#sc-8036; Santa Cruz Technology), HA-tag (#14793; Cell Signaling Technology), His-tag (#12698; Cell Signal Technology), FLAG-tag (#3724; Cell Signaling Technology), FLAG-tag (#8146; Cell Signaling Technology), ERK1/2 (#4695; Cell Signaling Technology), p-ERK1/2 T202/Y204 (#4370; Cell Signaling Technology), MEK1 (#9122; Cell Signaling Technology), p-MEK1 T292 (#07-852; Upstate, Darmstadt, Germany), BRAF (#sc-5284; Santa Cruz Technology), p-BRAF S445 (#2696; Cell Signaling Technology).

Techniques: Over Expression, Activation Assay, Immunofluorescence, Staining, Western Blot, Cytometry, In Vitro, Kinase Assay, Purification, Immunoprecipitation, Knock-Out

Journal: Cell Death Discovery

Article Title: Dual role of PID1 in regulating apoptosis induced by distinct anticancer-agents through AKT/Raf-1-dependent pathway in hepatocellular carcinoma

doi: 10.1038/s41420-023-01405-1

Figure Lengend Snippet: A Time-course analysis of p-Raf-1 S338 , p-Raf-1 S259 , p-MEK1 T292 , p-ERK1/2 T202/Y204 , BRAF and p-BRAF S445 in HepG2 cells with or without PID1 overexpression upon Sorafenib (10 μM) treatment. B Dynamics of Raf-1 activation and BRAF inactivation in HepG2 cells with or without PID1 overexpression upon Sorafenib (10 μM) treatment. p-Raf-1 S338 band density was normalized to actin and then normalized to t = 6 h. p-BRAF S445 band density was normalized to actin and then normalized to t = 0. C Endogenous interaction between Raf-1 and BRAF in HepG2 cells with or without PID1 overexpression upon Sorafenib (10 μM) treatment for 3 h or 6 h was examined by co-immunoprecipitation assay. D Western blot analysis of ERK1/2, p-ERK1/2 T202/Y204 , MEK1, p-MEK1 T292 , AKT, p-AKT T308 and β-actin in HepG2 cells with or without PID1 overexpression upon Sorafenib (10 μM) treatment for 3 h in the presence of L-779450 (20 μM). E HepG2 cells with or without PID1 overexpression upon Raf-1 knockdown were pretreated with Sorafenib (10 μM) for 3 h and then treated with insulin for 0.25 h. Western blot analysis of AKT, p-AKT T308 and β-actin was performed. F Cell apoptosis in HepG2 with or without PID1 overexpression combined with Raf-1 knockdown upon Sorafenib (10 μM) treatment for 24 h was examined by cytometry analysis. Data are expressed as mean ± SD ( n = 3). * p < 0.05; ** p < 0.01; ns not significant.

Article Snippet: Antibodies used to determine the protein expression were the following: PID1 (#27951; Signalway Antibody, Greenbelt, Maryland, USA), β-actin (#66009-1-lg; Proteintech, USA), Bcl-xL (#2764; Cell Signaling Technology, USA), Bcl2 (#12789-1-AP; Proteintech), BAD (#sc-8044 Santa Cruz Technology, Texas, USA), p-BAD S112 (#5284; Cell Signaling Technology), p-BAD S136 (#4366; Cell Signaling Technology), cytochrome c (#sc-13156; Santa Cruz Technology), cleaved caspase-3 (#9664; Cell Signaling Technology), Tom20 (#42406; Cell Signaling Technology), Raf-1 (#9422; Cell Signaling Technology), p-Raf-1 S259 (#9421; Cell Signaling Technology), p-Raf-1 S338 (#9427; Cell Signaling Technology), AKT (#4691; Cell Signaling Technology), p-AKT T308 (#13038; Cell Signaling Technology), HA-tag (#sc-7392; Santa Cruz Technology), His-tag (#sc-8036; Santa Cruz Technology), HA-tag (#14793; Cell Signaling Technology), His-tag (#12698; Cell Signal Technology), FLAG-tag (#3724; Cell Signaling Technology), FLAG-tag (#8146; Cell Signaling Technology), ERK1/2 (#4695; Cell Signaling Technology), p-ERK1/2 T202/Y204 (#4370; Cell Signaling Technology), MEK1 (#9122; Cell Signaling Technology), p-MEK1 T292 (#07-852; Upstate, Darmstadt, Germany), BRAF (#sc-5284; Santa Cruz Technology), p-BRAF S445 (#2696; Cell Signaling Technology).

Techniques: Over Expression, Activation Assay, Co-Immunoprecipitation Assay, Western Blot, Cytometry

Journal: Journal of Nanobiotechnology

Article Title: Extracellular vesicle-mediated gene therapy targets BRAF V600E -mutant colorectal cancer by inhibiting the MEK1/2-ERK1/2 pathway

doi: 10.1186/s12951-025-03205-4

Figure Lengend Snippet: Schematic illustration of the therapeutic system based on EVs used for the delivery of nucleic acid drugs targeting gene mutation sites for the precise treatment of CRC with the BRAF V600E mutation. siBRAF V600E was transfected into HEK293T cells via a lentiviral vector to stably produce EVs carrying siBRAF V600E . After treatment with these EVs- siBRAF V600E , the BRAF-MEK1/2-ERK1/2 pathway was inhibited in CRC cells. Subcutaneous xenograft tumor models and metastasis models of CRC cell lines were used to validate the therapeutic effects and toxicity of the EVs-siBRAF V600E . The efficacy of the EVs-siBRAF V600E was further confirmed via PDX models

Article Snippet: The primary antibodies used were against BRAF V600E (ZA-0668, ZSGB-BO, China), KI67 (27309-1-AP, Proteintech), BRAF (ab33899, Abcam), and phospho-p44/42 MAPK (p-ERK1/2) (4370s, Cell Signaling Technology).

Techniques: Mutagenesis, Transfection, Plasmid Preparation, Stable Transfection

Journal: Journal of Nanobiotechnology

Article Title: Extracellular vesicle-mediated gene therapy targets BRAF V600E -mutant colorectal cancer by inhibiting the MEK1/2-ERK1/2 pathway

doi: 10.1186/s12951-025-03205-4

Figure Lengend Snippet: BRAF mutation is a poor prognostic factor in CRC patients. ( A ) K-M survival curves of patients with BRAF mutation and wild-type CRC. ( B ) Multivariate COX regression model analysis of gender, stage and BRAF mutation status in CRC patients. ( C ) K-M survival curve of BRAF expression level in BRAF MT CRC. ( D ) Representative IHC images of BRAF V600E and KI-67 in CRC tissues. The scale bar represents 10 μm for the 10x objective and 5 μm for the 20x objective. ( E ) KI67 positivity rate in CRC tissues with BRAF V600E or BRAF WT . The data are reported as the means ± SDs of the experiments ( n = 3). Two-tailed Student’s t-tests, ** p < 0.01

Article Snippet: The primary antibodies used were against BRAF V600E (ZA-0668, ZSGB-BO, China), KI67 (27309-1-AP, Proteintech), BRAF (ab33899, Abcam), and phospho-p44/42 MAPK (p-ERK1/2) (4370s, Cell Signaling Technology).

Techniques: Mutagenesis, Expressing, Two Tailed Test

Journal: Journal of Nanobiotechnology

Article Title: Extracellular vesicle-mediated gene therapy targets BRAF V600E -mutant colorectal cancer by inhibiting the MEK1/2-ERK1/2 pathway

doi: 10.1186/s12951-025-03205-4

Figure Lengend Snippet: Construction and characterization of EVs-siBRAF V600E . ( A ) BRAF mRNA levels were analyzed by qRT-PCR. BRAF mRNA levels were measured after treating HEK293T, COLO320, and RKO cells with 50 µg/mL of EVs-siBRAF V600E for 24 h. ( B ) Construction of the therapeutic agent. ( C ) NTA analysis of EVs-NC (blue) and EVs-siBRAF V600E (red). ( D )( E ) TEM images of eVs-NC ( D ) and EVs-siBRAF V600E ( E ); scale bar: 200 nm. ( F ) WB analysis of the extracellular vesicle marker proteins, TSG101, CD81 and CD63. ( G ) Fluorescence microscopy images showing the uptake of EVs-NC and EVs-siBRAF V600E by RKO cells. The cell nuclei were stained with DAPI, and the EVs were stained with DiD; scale bar: 20 μm. ( H ) qRT-PCR analysis of siBRAF V600E levels in EVs-NC and EVs-siBRAF V600E . ( I ) WB analysis of BRAF protein levels. After COLO320 and RKO cells were treated with PBS, or 50 µg/mL of EVs-NC or EVs-siBRAF V600E for 24 h, cell proteins were extracted to measure BRAF protein expression. The data are reported as the means ± SDs of the experiments ( n = 3). Two-tailed Student’s t-tests for ( a ) and ( h ), and one-way ANOVA followed by Tukey test multiple comparisons for ( I ), * p < 0.05, ** p < 0.01 and *** p < 0.001

Article Snippet: The primary antibodies used were against BRAF V600E (ZA-0668, ZSGB-BO, China), KI67 (27309-1-AP, Proteintech), BRAF (ab33899, Abcam), and phospho-p44/42 MAPK (p-ERK1/2) (4370s, Cell Signaling Technology).

Techniques: Quantitative RT-PCR, Marker, Fluorescence, Microscopy, Staining, Expressing, Two Tailed Test

Journal: Journal of Nanobiotechnology

Article Title: Extracellular vesicle-mediated gene therapy targets BRAF V600E -mutant colorectal cancer by inhibiting the MEK1/2-ERK1/2 pathway

doi: 10.1186/s12951-025-03205-4

Figure Lengend Snippet: EVs-siBRAF V600E can inhibit BRAF V600E CRC cells. RKO ( A ) and HT29 ( B ) cells were treated with PBS, EVs-NC or EVs-siBRAF V600E for 48 h, followed by EdU assays to measure cell proliferation activity and imaging via fluorescence microscopy. Scale bar: 20 μm. RKO( C ) and HT29 ( D ) cells were treated with PBS, EVs-NC or EVs-siBRAF V600E for 5 days, and cell viability was measured via CCK-8 assays. ( E ) Colony formation. Images of single-cell-derived colonies formed by RKO and HT29 cells after treatment with PBS, EVs-NC or EVs-siBRAF V600E for 2 weeks. ( F ) Flow cytometry analysis of cell apoptosis. RKO and HT29 cells were treated with PBS, EVs-NC or EVs-siBRAF V600E for 24 h, stained with Annexin V-FITC and PI, and apoptosis was detected via flow cytometry. ( G ) Cell migration was assessed by wound healing assays. RKO cells were treated with PBS, EVs-NC or EVs-siBRAF V600E for 24 h. The concentration of EVs-NC or EVs-siBRAF V600E in all the treatments was 50 µg/mL. The data are reported as the means ± SDs of the experiments ( n = 3). Two-way ANOVA for ( A ) and ( B ), and one-way ANOVA followed by Tukey test multiple comparisons for the others, * p < 0.05, ** p < 0.01 and *** p < 0.001

Article Snippet: The primary antibodies used were against BRAF V600E (ZA-0668, ZSGB-BO, China), KI67 (27309-1-AP, Proteintech), BRAF (ab33899, Abcam), and phospho-p44/42 MAPK (p-ERK1/2) (4370s, Cell Signaling Technology).

Techniques: Activity Assay, Imaging, Fluorescence, Microscopy, CCK-8 Assay, Derivative Assay, Flow Cytometry, Staining, Migration, Concentration Assay

Journal: Journal of Nanobiotechnology

Article Title: Extracellular vesicle-mediated gene therapy targets BRAF V600E -mutant colorectal cancer by inhibiting the MEK1/2-ERK1/2 pathway

doi: 10.1186/s12951-025-03205-4

Figure Lengend Snippet: EVs-siBRAF V600E cannot inhibit BRAF WT CRC cells. COLO320( A ) and HCT116 ( B ) cells were treated with PBS, EVs-NC or EVs-siBRAF V600E for 5 days, and cell viability was measured via CCK-8 assays. COLO320( C ) and HCT116 ( D ) cells were treated with PBS, EVs-NC or EVs-siBRAF V600E for 48 h, followed by EdU assays to measure cell proliferation activity and imaging using fluorescence microscopy. Scale bar: 20 μm. ( E ) Colony formation. Images of single-cell-derived colonies formed by COLO320 and HCT116 cells after treatment with PBS, EVs-NC or EVs-siBRAF V600E for 2 weeks. ( F ) Flow cytometry analysis of cell apoptosis. COLO320 and HCT116 cells were treated with PBS, EVs-NC or EVs-siBRAF V600E for 24 h, stained with Annexin V-FITC and PI, and apoptosis was detected via flow cytometry. ( G ) Cell migration was assessed by wound healing assays. COLO320 and HCT116 cells were treated with PBS, EVs-NC or EVs-siBRAF V600E for 24 h. The concentration of EVs-NC and EVs-siBRAF V600E in all treatments was 50 µg/mL. The data are reported as the means ± SDs of the experiments ( n = 3). Two-way ANOVA for ( A ) and ( B ), and one-way ANOVA for the others

Article Snippet: The primary antibodies used were against BRAF V600E (ZA-0668, ZSGB-BO, China), KI67 (27309-1-AP, Proteintech), BRAF (ab33899, Abcam), and phospho-p44/42 MAPK (p-ERK1/2) (4370s, Cell Signaling Technology).

Techniques: CCK-8 Assay, Activity Assay, Imaging, Fluorescence, Microscopy, Derivative Assay, Flow Cytometry, Staining, Migration, Concentration Assay

Journal: Journal of Nanobiotechnology

Article Title: Extracellular vesicle-mediated gene therapy targets BRAF V600E -mutant colorectal cancer by inhibiting the MEK1/2-ERK1/2 pathway

doi: 10.1186/s12951-025-03205-4

Figure Lengend Snippet: EVs-siBRAF V600E Inhibit BRAF-MEK1/2-ERK1/2 Signaling Pathway in BRAF V600E CRC cells. ( A ) and ( B ) qRT-PCR analysis of BRAF transcription levels. RKO ( A ) and HT29 ( B ) cells were treated with PBS, EVs-NC or EVs-siBRAF V600E for 24 h, followed by detection of BRAF mRNA levels. ( C ) and ( D ) WB analysis of BRAF expression levels and MEK1/2-ERK1/2 phosphorylation levels. RKO ( C ) and HT29 ( D ) cells were treated with PBS, EVs-NC or EVs-siBRAF V600E for 24 h, and the BRAF protein levels and MEK1/2-ERK1/2 phosphorylation levels were measured. ( E ) and ( F ) qRT-PCR analysis of BRAF transcription levels. COLO320 ( E ) and HCT116 ( F ) cells were treated with PBS, EVs-NC or EVs-siBRAF V600E for 24 h, followed by the detection of BRAF mRNA levels. ( G ) and (H) WB analysis of BRAF expression levels and MEK1/2-ERK1/2 phosphorylation levels. COLO320 ( G ) and HT29 ( H ) cells were treated with PBS, EVs-NC or EVs-siBRAF V600E for 24 h, and the BRAF protein levels and MEK1/2-ERK1/2 phosphorylation levels were measured. The concentration of EVs-NC and EVs-siBRAF V600E in all the treatments was 50 µg/mL. ( I ) Schematic illustration of the mechanism by which EVs-siBRAF V600E treat CRC cells. The data are reported as the means ± SDs of the experiments ( n = 3). One-way ANOVA followed by Tukey test multiple comparisons, * p < 0.05, ** p < 0.01 and *** p < 0.001

Article Snippet: The primary antibodies used were against BRAF V600E (ZA-0668, ZSGB-BO, China), KI67 (27309-1-AP, Proteintech), BRAF (ab33899, Abcam), and phospho-p44/42 MAPK (p-ERK1/2) (4370s, Cell Signaling Technology).

Techniques: Quantitative RT-PCR, Expressing, Phospho-proteomics, Concentration Assay

Journal: Journal of Nanobiotechnology

Article Title: Extracellular vesicle-mediated gene therapy targets BRAF V600E -mutant colorectal cancer by inhibiting the MEK1/2-ERK1/2 pathway

doi: 10.1186/s12951-025-03205-4

Figure Lengend Snippet: EVs-siBRAF V600E inhibit BRAF V600E CRC in vivo. ( A ) Animal model handling protocol. ( B ) Subcutaneous tumor volume changes. Tumor volume measurements were performed every 3 days. ( C ) Images of the subcutaneous tumors in the three groups. ( D ) The weights of the subcutaneous tumors. ( E ) Immunohistochemistry images of BRAF and p-ERK1/2. Scale bar: 50 μm. ( F )~( I ) WB detection of BRAF expression and MEK1/2-ERK1/2 phosphorylation levels in subcutaneous tumor tissues. ( J ) Bioluminescence images of lung metastases. ( K ) Bioluminescence images of liver metastases. Each group of animals was treated with PBS, EVs-NC or EVs-siBRAF V600E . Each mouse was injected with 20 µg of extracellular vesicles per injection. Subcutaneous tumors: n = 5 per group. Lung metastasis and liver metastasis: n = 4 per group. The data are reported as the means ± SDs of the experiments. Two-way ANOVA for ( B ), and one-way ANOVA followed by Tukey test multiple comparisons for the others, * p < 0.05, ** p < 0.01 and *** p < 0.001

Article Snippet: The primary antibodies used were against BRAF V600E (ZA-0668, ZSGB-BO, China), KI67 (27309-1-AP, Proteintech), BRAF (ab33899, Abcam), and phospho-p44/42 MAPK (p-ERK1/2) (4370s, Cell Signaling Technology).

Techniques: In Vivo, Animal Model, Immunohistochemistry, Expressing, Phospho-proteomics, Injection

Journal: Journal of Nanobiotechnology

Article Title: Extracellular vesicle-mediated gene therapy targets BRAF V600E -mutant colorectal cancer by inhibiting the MEK1/2-ERK1/2 pathway

doi: 10.1186/s12951-025-03205-4

Figure Lengend Snippet: Efficacy of EVs-siBRAF V600E in human CRC PDX models. ( A ) PDX model handling protocol. ( B ) Subcutaneous tumor volume changes in the BRAF V600E PDX models. Tumor volume measurements were performed every 3 days. ( C )Images of subcutaneous tumors from the BRAF V600E PDX models. ( D ) Subcutaneous tumor weights of the BRAF V600E PDX models. ( E ) Immunohistochemistry images of BRAF and p-ERK1/2 in the BRAF V600E PDX models. Scale bar: 50 μm. ( F ) and ( G ) Subcutaneous tumor volume changes in the BRAF WT PDX models. Tumor volumes of BRAF WT PDX1( F ) and PDX2( G ) were measured every 3 days. ( H ) Images of subcutaneous tumors from the BRAF WT PDX1 models. ( I ) Subcutaneous tumor weights of the BRAF WT PDX1 models. ( J ) Images of subcutaneous tumors from the BRAF WT PDX2 models. ( K ) Subcutaneous tumor weights of the BRAF WT PDX2 models. Each group of animals was treated with PBS, EVs-NC or EVs-siBRAF V600E . Each mouse was injected with 20 µg of extracellular vesicles per injection. N = 6 for each group of BRAF V600E PDX models. N = 5 for each group in the BRAF WT PDX1 and PDX2 models. The data are reported as the means ± SDs of the experiments. Two-way ANOVA for ( B ), ( F ) and ( G ), and one-way ANOVA followed by Tukey test multiple comparisons for the others, * p < 0.05, ** p < 0.01 and *** p < 0.001

Article Snippet: The primary antibodies used were against BRAF V600E (ZA-0668, ZSGB-BO, China), KI67 (27309-1-AP, Proteintech), BRAF (ab33899, Abcam), and phospho-p44/42 MAPK (p-ERK1/2) (4370s, Cell Signaling Technology).

Techniques: Immunohistochemistry, Injection

Journal: Journal of Nanobiotechnology

Article Title: Extracellular vesicle-mediated gene therapy targets BRAF V600E -mutant colorectal cancer by inhibiting the MEK1/2-ERK1/2 pathway

doi: 10.1186/s12951-025-03205-4

Figure Lengend Snippet: Tumor targeting capability and safety of EVs-siBRAF V600E in vivo. ( A ) Biodistribution images of EVs-NC or EVs-siBRAF V600E in major organs and tumors. ( B ) Semi-quantitative analysis of the distribution in tumors. ( C )~( D ) After treatment with PBS, EVs-NC or EVs-siBRAF V600E , related serum biochemistry indicators were analyzed, including hepatic function enzymes, such as ALT ( C ) and AST ( D ); renal function enzymes, such as BUN ( E ) and CREA ( F ); and cardiac function indicators, such as CK ( G ) and CK-MB ( H ), were analyzed in RKO xenograft models. ( I ) ~( L ) Representative images of histological assessments of major organs. After treatment with PBS, EVs-NC and EVs-siBRAF V600E , the lungs, kidneys, spleens and livers of RKO xenograft models from each group were collected. The data are reported as the means ± SDs of the experiments ( n = 5). One-way ANOVA

Article Snippet: The primary antibodies used were against BRAF V600E (ZA-0668, ZSGB-BO, China), KI67 (27309-1-AP, Proteintech), BRAF (ab33899, Abcam), and phospho-p44/42 MAPK (p-ERK1/2) (4370s, Cell Signaling Technology).

Techniques: In Vivo