braf v600e antibody clone ve1  (Spring Bioscience)

Journal: Journal for immunotherapy of cancer

Article Title: Exploring markers of immunoresponsiveness in papillary thyroid carcinoma and future treatment strategies.

doi: 10.1136/jitc-2023-008505

Figure Lengend Snippet: Figure 3 Expression of immunosuppressive markers correlates to BRAF mutation. (A) The tumor tissues of 19 PTC cases were stained with CD274 antibody, and the expression intensity was scored by a COH pathologist. Grade 1 represents the lowest expression, and grade 3+ represents the highest. (B) The tumor tissue was processed to detect CD73, and the expression intensity was scored where grade 1 represents the lowest expression and grade 3+ represents the highest. (C) The images represent the cells positive for cancer cells (TTF-brown), T cells CD8 (blue) and CD4 (pink) in the two representative tumor tissue of BRAF-mutated and WT samples. (D) The plot illustrates the percentage of lymphocyte infiltration within the tumor area, as assessed by the pathologist. The lymphocytic infiltration ratio is notably higher for mutant patients (red dots) compared with wild-type patients (blue). Additionally, the percentages of CD4 and CD8 lymphocytes were analyzed. The findings suggest a higher contribution of CD4 lymphocytes compared with CD8, reinforcing the cell type infiltration data. PTC, papillary thyroid cancer; WT, wild-type. * indicates a p value of <0.05

Article Snippet: Antibodies against BRAF (cat #: 14814), CD274 (cat #: 60475), CD276 (cat #:58798), CD73 (cat # 13160), RAS (cat #: 91054), CMTM6 (cat #: 19130), phospho- Rb (S807/811) (cat#:8516), KRAS (cat #: 33197), p- ERK (cat #: 4370) were purchased from Cell Signaling Technology (Danvers, Massachusetts, USA).

Techniques: Expressing, Mutagenesis, Staining

Journal: Journal for immunotherapy of cancer

Article Title: Exploring markers of immunoresponsiveness in papillary thyroid carcinoma and future treatment strategies.

doi: 10.1136/jitc-2023-008505

Figure Lengend Snippet: Figure 4 BRAF knockdown induced CD274 expression. (A) The RNA extracted from four PTC cell lines was used to determine the expression of BRAF, CD274, CD73, CD200, CD276, ENTPD1, and CD200. The expression in the mutant was compared with the WT cell type. (B) The immunoblot represents the expression of proteins in the BRAF-mutated and WT cell lines. (C) The bar graph represents the changes in the mRNA expression of CD274 and other immunosuppressive markers on knocking down BRAF. (D) Immunoblot represents the knockdown of BRAF and upregulation of CD274 in the BRAF-mutated cell lines T32 and T85 (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ANOVA test). ANOVA, analysis of variance; PTC, papillary thyroid cancer; WT, wild-type.

Article Snippet: Antibodies against BRAF (cat #: 14814), CD274 (cat #: 60475), CD276 (cat #:58798), CD73 (cat # 13160), RAS (cat #: 91054), CMTM6 (cat #: 19130), phospho- Rb (S807/811) (cat#:8516), KRAS (cat #: 33197), p- ERK (cat #: 4370) were purchased from Cell Signaling Technology (Danvers, Massachusetts, USA).

Techniques: Knockdown, Expressing, Mutagenesis, Western Blot

Journal: Journal for immunotherapy of cancer

Article Title: Exploring markers of immunoresponsiveness in papillary thyroid carcinoma and future treatment strategies.

doi: 10.1136/jitc-2023-008505

Figure Lengend Snippet: Figure 5 Cell cycle arrest correlates with CD274 expression. (A, B) The T85 cell line was treated with increasing concentration of BRAF inhibitors, and changes in the proliferation were measured using the live cell imaging system. (C, D) Data representing the proliferation changes in the T68 cell lines with respect to BRAF inhibitor treatment. (E) FACS data representing the changes in the cell cycle stages of T68 with respect to BRAF inhibitors where dabrafenib induced stronger cell cycle arrest. (F) The qPCR data represents the changes in the expression of BRAF, CD274 and CD73 in the T68 and T85 cells with respect to the BRAF inhibitor treatment. (G) The immunoblot represents the changes in the CD274 and CD73 expression on combining dabrafenib with either binimetinib, temsirolimus, or abemaciclib (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ANOVA test). ANOVA, analysis of variance.

Article Snippet: Antibodies against BRAF (cat #: 14814), CD274 (cat #: 60475), CD276 (cat #:58798), CD73 (cat # 13160), RAS (cat #: 91054), CMTM6 (cat #: 19130), phospho- Rb (S807/811) (cat#:8516), KRAS (cat #: 33197), p- ERK (cat #: 4370) were purchased from Cell Signaling Technology (Danvers, Massachusetts, USA).

Techniques: Expressing, Concentration Assay, Live Cell Imaging, Western Blot

Journal: Journal for immunotherapy of cancer

Article Title: Exploring markers of immunoresponsiveness in papillary thyroid carcinoma and future treatment strategies.

doi: 10.1136/jitc-2023-008505

Figure Lengend Snippet: Figure 6 BRAF inhibitor-induced transcriptional changes upstream of CD274. (A) Cartoon showing the design of CD274- promoter-luciferase constructs used for determining the changes in transcriptional activity postdrug treatment. (B) The box plot representing the luciferase activity determined in the cells post 48 hours of transfection, where 2 Kb promoter sequence correlated with maximum activity. (C) Luciferase activity measured in the cells post 48 hours of BRAF or KRAS knockdown. (D) The bar graph represents the difference in the promoter activity between the T85 and T32 cell lines. (E, F) Luciferase (Promoter) activity correlated with increased BRAF inhibitor concentration in the T68 and T85 cells. (****p<0.0001, ANOVA test). ANOVA, analysis of variance.

Article Snippet: Antibodies against BRAF (cat #: 14814), CD274 (cat #: 60475), CD276 (cat #:58798), CD73 (cat # 13160), RAS (cat #: 91054), CMTM6 (cat #: 19130), phospho- Rb (S807/811) (cat#:8516), KRAS (cat #: 33197), p- ERK (cat #: 4370) were purchased from Cell Signaling Technology (Danvers, Massachusetts, USA).

Techniques: Luciferase, Construct, Activity Assay, Transfection, Sequencing, Knockdown, Concentration Assay

Journal: Journal for immunotherapy of cancer

Article Title: Exploring markers of immunoresponsiveness in papillary thyroid carcinoma and future treatment strategies.

doi: 10.1136/jitc-2023-008505

Figure Lengend Snippet: Figure 8 The synergistic effect of BRAF and checkpoint inhibitors in immune cell-mediated killing. (A) The line graph represents the growth kinetics of T85 cells when cultured in combination with activated CD8 cells. The bar graph represents the comparison in growth between monoculture and cocultured T85 cells at 0 and 72 hours time points. (B) The growth kinetics of T32 cells when cultured in combination with activated CD8 cells. (C) The images represent the effect of dabrafenib, CD8, atezolizumab, and IL-2 on the T85 (GFP) cell growth. (D) The growth kinetics of T85 cells when cultured in combination with activated CD8 cells, IL-2, and atezolizumab. (E) The growth kinetics of T85 cells, when cultured in combination with dabrafenib, activated CD8 cells, IL-2 and atezolizumab. (F) The images represent the effect of CD8, atezolizumab, and IL-2 on the T85 (GFP) derived spheroids. Loss of GFP represents a loss of spheroid viability. (G) The bar graph represents the changes in the spheroid viability when cocultured with CD8, IL-2, and atezolizumab. (H) The changes in the spheroid viability when cocultured with dabrafenib, CD8, IL-2, and atezolizumab. (I) The immunoblot represents the changes in the CD274 expression in T85 cells on exposure to IL-2 and dabrafenib n monoculture or coculture. **** indicates P value < 0.001.

Article Snippet: Antibodies against BRAF (cat #: 14814), CD274 (cat #: 60475), CD276 (cat #:58798), CD73 (cat # 13160), RAS (cat #: 91054), CMTM6 (cat #: 19130), phospho- Rb (S807/811) (cat#:8516), KRAS (cat #: 33197), p- ERK (cat #: 4370) were purchased from Cell Signaling Technology (Danvers, Massachusetts, USA).

Techniques: Cell Culture, Comparison, Derivative Assay, Western Blot, Expressing

Journal: Cancers

Article Title: Anti-MAPK Targeted Therapy for Ameloblastoma: Case Report with a Systematic Review

doi: 10.3390/cancers16122174

Figure Lengend Snippet: Demographics (M male, F female, NS non-specified, IHC Immunohistochemistry, NGS next-generation sequencing, PCR polymerase chain reaction, BID twice a day, QD once a day, CR complete response, NCR near-complete response, PR partial response, SD stable disease, N no, TTT treatment).

Article Snippet: The histopathologic examination revealed ameloblastoma ( A), with positive immunohistostaining for BRAF V600E (clone VE1, Ventana Medical Systems, B).

Techniques: Immunohistochemistry, Next-Generation Sequencing, Polymerase Chain Reaction