Journal: bioRxiv

Article Title: Crosstalk in skin: Loss of desmoglein 1 in keratinocytes inhibits BRAF V600E -induced cellular senescence in human melanocytes

doi: 10.1101/2023.02.16.528886

Figure Lengend Snippet: A , Gene set enrichment analysis compared differentially expressed gene sets to published signatures of melanoma/melanocyte signaling. NES, normalized enrichment score. B , Volcano plots revealed log2 fold change against nominal p values (-log10) from differential expression analysis of mRNA sequencing of melanocytes with different BRAF statuses (control, WT BRAF and BRAF V600E ) and treated with control or Dsg1 knockdown conditioned media. Select genes involved in differentiation, pigmentation, and proliferation are marked.

Article Snippet: The following primary antibodies were used: BRAF (#9433), p44/42 MAPK (#9107), P-p44/42 MAPK (#4370), and p16 (#80772) from Cell Signaling Technology, BRAF V600E (ab200535) from Abcam, Dsg1 (#32-6000) from Thermo Fisher Scientific, GAPDH (G9545) from MilliporeSigma, and Tubulin (12G10) from Developmental Studies Hybridoma Bank.

Techniques: Expressing, Sequencing

Journal: bioRxiv

Article Title: Crosstalk in skin: Loss of desmoglein 1 in keratinocytes inhibits BRAF V600E -induced cellular senescence in human melanocytes

doi: 10.1101/2023.02.16.528886

Figure Lengend Snippet: Human primary melanocytes were transduced with WT BRAF or BRAF V600E lentivirus, control or Dsg1-deficient CM were added 24 h after transduction and cells continued to be cultured in CM before they were harvested. A , Relative cell numbers over time. B , EdU flow cytometry assay to determine cell proliferation. The figures showed a clear separation of proliferating cells which have incorporated EdU in control and WT BRAF melanocytes but not in BRAF V600E melanocytes. C , Quantification of EdU flow cytometry assay. D , Annexin V/dead cell apoptosis flow cytometry assay was performed to detect cell death. E , Quantification of cell death by flow cytometry assay. F , Senescence-associated β-galactosidase activity was detected by flow cytometry assay. Right shift of the peak indicated the increase in senescence. G , Quantification of senescence flow cytometry assay. H , Expression of the indicated proteins was determined by Western blot. Results were reported as mean ± SD (n=3), statistical analysis was performed using two-way ANOVA with multiple comparisons (*, P < 0.05; ***, P < 0.001).

Article Snippet: The following primary antibodies were used: BRAF (#9433), p44/42 MAPK (#9107), P-p44/42 MAPK (#4370), and p16 (#80772) from Cell Signaling Technology, BRAF V600E (ab200535) from Abcam, Dsg1 (#32-6000) from Thermo Fisher Scientific, GAPDH (G9545) from MilliporeSigma, and Tubulin (12G10) from Developmental Studies Hybridoma Bank.

Techniques: Transduction, Cell Culture, Flow Cytometry, Activity Assay, Expressing, Western Blot

Journal: bioRxiv

Article Title: Crosstalk in skin: Loss of desmoglein 1 in keratinocytes inhibits BRAF V600E -induced cellular senescence in human melanocytes

doi: 10.1101/2023.02.16.528886

Figure Lengend Snippet: A , Bar graph of top enriched pathways represented in genes that were significantly up/down-regulated by Dsg1 knockdown CM. B , Heatmap with hierarchical clustering revealed the expression of the top 50 differentially expressed genes between BRAF V600E melanocytes treated with control and Dsg1-deficient CM.

Article Snippet: The following primary antibodies were used: BRAF (#9433), p44/42 MAPK (#9107), P-p44/42 MAPK (#4370), and p16 (#80772) from Cell Signaling Technology, BRAF V600E (ab200535) from Abcam, Dsg1 (#32-6000) from Thermo Fisher Scientific, GAPDH (G9545) from MilliporeSigma, and Tubulin (12G10) from Developmental Studies Hybridoma Bank.

Techniques: Expressing

Journal: bioRxiv

Article Title: Crosstalk in skin: Loss of desmoglein 1 in keratinocytes inhibits BRAF V600E -induced cellular senescence in human melanocytes

doi: 10.1101/2023.02.16.528886

Figure Lengend Snippet: A , qRT-PCR was performed to confirm the up-regulation of NTN4 expression in BRAF V600E -transduced cells treated with Dsg1 knockdown CM. Mean ± SD depicted (n=3), statistical analysis was performed using Student’s t-test (**, p<0.01). B , Immunofluorescence analysis confirmed higher expression of Netrin-4 protein upon Dsg1-deficient CM treatment. C and D , qRT-PCR and immunofluorescence staining were carried out to confirm the knockdown of NTN4 expression (both mRNA and protein) by shRNAs. Results in C was reported as Mean ± SD (n=3), statistical analysis was performed using one-way ANOVA with multiple comparisons (***, p<0.001). E , Senescence-associated β-galactosidase expression was detected by X-gal staining assay. F , Quantification of X-gal staining assay. Mean ± SD depicted (n=3), statistical analysis was performed using two-way ANOVA with multiple comparisons (**, p<0.01).

Article Snippet: The following primary antibodies were used: BRAF (#9433), p44/42 MAPK (#9107), P-p44/42 MAPK (#4370), and p16 (#80772) from Cell Signaling Technology, BRAF V600E (ab200535) from Abcam, Dsg1 (#32-6000) from Thermo Fisher Scientific, GAPDH (G9545) from MilliporeSigma, and Tubulin (12G10) from Developmental Studies Hybridoma Bank.

Techniques: Quantitative RT-PCR, Expressing, Immunofluorescence, Staining

Journal: Frontiers in Endocrinology

Article Title: The high degree of similarity in histopathological and clinical characteristics between radiogenic and sporadic papillary thyroid microcarcinomas in young patients

doi: 10.3389/fendo.2022.970682

Figure Lengend Snippet: Characteristics of the radiogenic and sporadic papillary thyroid microcarcinomas in patients aged up to 30 years.

Article Snippet: In brief, a mouse monoclonal anti-BRAF (mutated V600E) antibody (VE1) ab228461(Abcam) at a 1:100 dilution and the Novolink Polymer Detection System (250T) (Leica RE7140-K) were used to detect IHC reaction product.

Techniques: Labeling, Dissection, Activity Assay

Journal: Frontiers in Endocrinology

Article Title: The high degree of similarity in histopathological and clinical characteristics between radiogenic and sporadic papillary thyroid microcarcinomas in young patients

doi: 10.3389/fendo.2022.970682

Figure Lengend Snippet: Characteristics of the radiogenic and sporadic papillary thyroid microcarcinomas in pediatric patients aged ≤ 18 years.

Article Snippet: In brief, a mouse monoclonal anti-BRAF (mutated V600E) antibody (VE1) ab228461(Abcam) at a 1:100 dilution and the Novolink Polymer Detection System (250T) (Leica RE7140-K) were used to detect IHC reaction product.

Techniques: Labeling, Dissection, Activity Assay

Journal: Frontiers in Endocrinology

Article Title: The high degree of similarity in histopathological and clinical characteristics between radiogenic and sporadic papillary thyroid microcarcinomas in young patients

doi: 10.3389/fendo.2022.970682

Figure Lengend Snippet: Characteristics of the radiogenic and sporadic papillary thyroid microcarcinomas in young adult patients aged 19-30 years.

Article Snippet: In brief, a mouse monoclonal anti-BRAF (mutated V600E) antibody (VE1) ab228461(Abcam) at a 1:100 dilution and the Novolink Polymer Detection System (250T) (Leica RE7140-K) were used to detect IHC reaction product.

Techniques: Labeling, Dissection, Activity Assay

Journal: Frontiers in Endocrinology

Article Title: The high degree of similarity in histopathological and clinical characteristics between radiogenic and sporadic papillary thyroid microcarcinomas in young patients

doi: 10.3389/fendo.2022.970682

Figure Lengend Snippet: Characteristics of the radiogenic and sporadic papillary thyroid microcarcinomas sized up to 5 mm.

Article Snippet: In brief, a mouse monoclonal anti-BRAF (mutated V600E) antibody (VE1) ab228461(Abcam) at a 1:100 dilution and the Novolink Polymer Detection System (250T) (Leica RE7140-K) were used to detect IHC reaction product.

Techniques: Labeling, Dissection, Activity Assay

Journal: Frontiers in Endocrinology

Article Title: The high degree of similarity in histopathological and clinical characteristics between radiogenic and sporadic papillary thyroid microcarcinomas in young patients

doi: 10.3389/fendo.2022.970682

Figure Lengend Snippet: Sporadic recurrent MPTC sized 8 mm (pT1aN0M0 at the first operation) removed from a 22-year-old male patient: (A–D) primary tumor, (E–H) the RAI-R recurrent metastasis. (A) Nonencapsulated primary tumor with follicular-papillary (conventional) growth pattern with extrathyroidal extension to the connective tissue, H&E, 15X magnification. (B) Fragment of the primary tumor with papillary structures featuring tall cell areas and oncocytic changes, H&E, 400X magnification. (C) Primary tumor: positive IHC reaction with the anti-BRAF (mutated V600E) antibody, 400X magnification. (D) Primary tumor: IHC reaction with Ki67 (Clone MIB-1) antibody (Ki67 LI 3.1%), 400X magnification. (E) RAI-R recurrent metastasis with cystic changes removed 1.3 years after the first surgery, H&E, 15X magnification. (F) Fragment of the RAI-R recurrent metastasis with solid structure and oncocytic changes, H&E, 400X magnification. (G) fragment of the RAI-R recurrent metastasis: positive IHC reaction with anti-BRAF (mutated V600E) antibody, 400X magnification. (H) RAI-R recurrent metastasis: IHC reaction with Ki67 (Clone MIB-1) antibody (Ki67 LI 3.3%), 400X magnification.

Article Snippet: In brief, a mouse monoclonal anti-BRAF (mutated V600E) antibody (VE1) ab228461(Abcam) at a 1:100 dilution and the Novolink Polymer Detection System (250T) (Leica RE7140-K) were used to detect IHC reaction product.

Techniques:

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