Journal: International Journal of Neuropsychopharmacology

Article Title: DHF-7 Ameliorates Behavioral Disorders and White Matter Lesions by Regulating BDNF and Fyn in a Mouse Model of Schizophrenia Induced by Cuprizone and MK-801

doi: 10.1093/ijnp/pyac022

Figure Lengend Snippet: Effects of DHF-7 on the expression of myelin-associated proteins in the corpus callosum of the mouse model induced by cuprizone and MK-801. (A) Representative images of Myelin basic protein (MBP) immunohistochemistry staining. Scale bar = 50 μm. (B) Optical density of MBP immunohistochemistry. The optical density in the control group is taken as 100%, n = 4. (C) Representative western-blot images of MBP and 2’,3’-Cyclic nucleotide-3’-phosphodiesterase (CNP). (D, E) Quantitative analysis of MBP and CNPase expression from the western-blot images, respectively. The ratio of MBP and CNPase to β-actin in the control group was taken as 100%, n = 3. Data are expressed as mean ± SEM. ## P < .01, model group vs. control group; * P < .05, ** P < .01, drug groups vs. model group.

Article Snippet: The following primary antibodies were used in this study: rat anti-MBP (Merck Millipore), rabbit anti-2’,3’-Cyclic nucleotide-3’-phosphodiesterase (CNPase) (Merck Millipore), rabbit anti- brain-derived neurotrophic factor (BDNF) (Abcam), rabbit anti-tyrosine kinase receptor B (TrkB; Abcam), rabbit anti-p-TrkB (phospho Tyr-705, Abcam), rabbit anti-Fyn (Abcam), rabbit anti-Fyn (phospho-Tyr-416; Abcam), rabbit anti-NMDA receptor subunit 2 B (NMDAR2B; Abcam), rabbit anti-NMDAR2B (phospho-Tyr-1472) (Abcam), rabbit anti-Braf, rabbit anti-p-Braf (Ser-445), rabbit anti-MEK1/2, rabbit anti-p-MEK1/2 (Ser217/221), rabbit anti-ERK1/2, and rabbit anti-p-ERK1/2 (Thr202/Try204) from Cell Signaling Technology, USA, and mouse anti-β-actin (Sigma-Aldrich).

Techniques: Expressing, Immunohistochemistry, Staining, Control, Western Blot

Journal: EMBO Molecular Medicine

Article Title: Intra- and inter-tumor heterogeneity in a vemurafenib-resistant melanoma patient and derived xenografts

doi: 10.15252/emmm.201404914

Figure Lengend Snippet: Hematoxylin–eosin (H&E) and p-ERK stainings on FFPE material of all metastases showed that all vemurafenib-resistant tumors had reactivation of the MAPK pathway. Scale bar represents 100 μm. CNA profiles were generated from the WES data with germ line DNA as a reference. Colors represent segmented log2 ratio values with red for gain and blue for loss. Further inspection of chromosome 7 where BRAF is located revealed amplification (7q34) of this region in three vemurafenib-resistant metastases, namely M032R1, M032R2, and M032R5. qPCR was performed on gDNA retrieved from each of the metastases, using primers for BRAF and CRAF and normalized on LINE levels. Bars represent the mean of three replicates, error bars indicate standard deviation. The results confirmed that BRAF was amplified in M032R1, M032R2, and M032R5. Staining for BRAF V600E with a mutant epitope-specific antibody confirmed the upregulation of BRAF V600E in R1, R2, and R5. Scale bar represents 100 μm.

Article Snippet: The following antibodies were used: mouse anti-p-ERK1/2 (E10, #9106), rabbit anti-ERK1/2 (9102), rabbit anti-p-MEK (41G9, #9154), mouse anti-MEK (L38C12, #4649) from Cell Signaling, mouse anti-BRAF V600E (VE1, Spring Bioscience), rabbit anti-p-RSK (04-419, Millipore), rabbit anti-RSK (9355, Cell Signaling), mouse anti-B-RAF (F7, Santa Cruz), mouse anti-vinculin (V9131, Sigma), and rabbit anti-p-AKT (D9E, #4060, Cell Signaling).

Techniques: Generated, Amplification, Standard Deviation, Staining, Mutagenesis

Journal: EMBO Molecular Medicine

Article Title: Intra- and inter-tumor heterogeneity in a vemurafenib-resistant melanoma patient and derived xenografts

doi: 10.15252/emmm.201404914

Figure Lengend Snippet: Immunoblotting for MAPK pathway components confirmed reactivation of p-ERK and p-MEK in all resistant metastases. Immunoblotting for M032 patient’s samples using an antibody directed against either the mutant BRAF V600E epitope or an N-terminal region of BRAF revealed the expression of a shorter variant of BRAF V600E in M032R3, of approximately 80 kDa. This 80-kDa band could not be detected with the N-terminus-recognizing antibody. Source data are available online for this figure.

Article Snippet: The following antibodies were used: mouse anti-p-ERK1/2 (E10, #9106), rabbit anti-ERK1/2 (9102), rabbit anti-p-MEK (41G9, #9154), mouse anti-MEK (L38C12, #4649) from Cell Signaling, mouse anti-BRAF V600E (VE1, Spring Bioscience), rabbit anti-p-RSK (04-419, Millipore), rabbit anti-RSK (9355, Cell Signaling), mouse anti-B-RAF (F7, Santa Cruz), mouse anti-vinculin (V9131, Sigma), and rabbit anti-p-AKT (D9E, #4060, Cell Signaling).

Techniques: Western Blot, Mutagenesis, Expressing, Variant Assay

Journal: EMBO Molecular Medicine

Article Title: Intra- and inter-tumor heterogeneity in a vemurafenib-resistant melanoma patient and derived xenografts

doi: 10.15252/emmm.201404914

Figure Lengend Snippet: Resistance mechanisms discovered in each of the lesions have been identified: a) in patients samples by WES ( BRAF V600E amplification in M032R1, R2 and R5; MEK1 T55delinsRT in M032R4 and M032) and immunoblotting (of alternative form of BRAF in M032R3); and b) in PDX by IHC and qPCR ( BRAF V600E amplification in M032R2 and R5) and PCR analysis ( MEK1 T55delinsRT in M032R1 and R4). Gray arrows indicate that the lesion was at the back side. ampl., amplification; alt., alternative.

Article Snippet: The following antibodies were used: mouse anti-p-ERK1/2 (E10, #9106), rabbit anti-ERK1/2 (9102), rabbit anti-p-MEK (41G9, #9154), mouse anti-MEK (L38C12, #4649) from Cell Signaling, mouse anti-BRAF V600E (VE1, Spring Bioscience), rabbit anti-p-RSK (04-419, Millipore), rabbit anti-RSK (9355, Cell Signaling), mouse anti-B-RAF (F7, Santa Cruz), mouse anti-vinculin (V9131, Sigma), and rabbit anti-p-AKT (D9E, #4060, Cell Signaling).

Techniques: Amplification, Western Blot

Journal: BMC Cancer

Article Title: microRNA-193a-3p is specifically down-regulated and acts as a tumor suppressor in BRAF -mutated colorectal cancer

doi: 10.1186/s12885-017-3739-x

Figure Lengend Snippet: Clinical characteristics between a screening and a validation set of patients

Article Snippet: Anti-BRAF rabbit monoclonal antibody (#9433, Cell Signaling Technology, Danvers, MA, USA), anti-p-BRAF (Ser445) rabbit monoclonal antibody (#2696, Cell Signaling Technology), anti-p-MEK1/2 (Ser217/221) rabbit polyclonal antibody (#9121, Cell Signaling Technology), anti-p-ERK1/2 (Ser217/221) rabbit monoclonal antibody (#4094, Cell Signaling Technology), and anti-α-tubulin mouse monoclonal antibody (Sigma-Aldrich) were used as primary antibodies for detection of the specific proteins.

Techniques: Biomarker Discovery

Journal: BMC Cancer

Article Title: microRNA-193a-3p is specifically down-regulated and acts as a tumor suppressor in BRAF -mutated colorectal cancer

doi: 10.1186/s12885-017-3739-x

Figure Lengend Snippet: Screening of candidate miRNAs, which were significantly altered in BRAF -mutant colorectal cancers ( n = 15) compared to KRAS/BRAF -wild-type colorectal cancers (n = 15), using a genome-wide miRNA expression analysis. a The top five dysregulated miRNAs from the miRNA microarray analysis. b The results of the microarray analysis were technically validated using Taqman real-time RT-PCR. Mann–Whitney U test was used to analyze statistical differences

Article Snippet: Anti-BRAF rabbit monoclonal antibody (#9433, Cell Signaling Technology, Danvers, MA, USA), anti-p-BRAF (Ser445) rabbit monoclonal antibody (#2696, Cell Signaling Technology), anti-p-MEK1/2 (Ser217/221) rabbit polyclonal antibody (#9121, Cell Signaling Technology), anti-p-ERK1/2 (Ser217/221) rabbit monoclonal antibody (#4094, Cell Signaling Technology), and anti-α-tubulin mouse monoclonal antibody (Sigma-Aldrich) were used as primary antibodies for detection of the specific proteins.

Techniques: Mutagenesis, Genome Wide, Expressing, Microarray, Quantitative RT-PCR, MANN-WHITNEY

Journal: BMC Cancer

Article Title: microRNA-193a-3p is specifically down-regulated and acts as a tumor suppressor in BRAF -mutated colorectal cancer

doi: 10.1186/s12885-017-3739-x

Figure Lengend Snippet: Validation of the miRNAs dysregulated in BRAF -mutant tumors, in another set of patients with colorectal cancer. The expression levels of the five miRNAs were validated in a different set including KRAS/BRAF -wild-type ( n = 30) and BRAF -mutant colorectal cancers ( n = 4). KRAS -mutant cancers ( n = 20) and adjacent normal mucosa ( n = 11) were additionally analyzed for the five miRNA expression. Mann-Whitney U test was used to analyze statistical differences

Article Snippet: Anti-BRAF rabbit monoclonal antibody (#9433, Cell Signaling Technology, Danvers, MA, USA), anti-p-BRAF (Ser445) rabbit monoclonal antibody (#2696, Cell Signaling Technology), anti-p-MEK1/2 (Ser217/221) rabbit polyclonal antibody (#9121, Cell Signaling Technology), anti-p-ERK1/2 (Ser217/221) rabbit monoclonal antibody (#4094, Cell Signaling Technology), and anti-α-tubulin mouse monoclonal antibody (Sigma-Aldrich) were used as primary antibodies for detection of the specific proteins.

Techniques: Biomarker Discovery, Mutagenesis, Expressing, MANN-WHITNEY

Journal: BMC Cancer

Article Title: microRNA-193a-3p is specifically down-regulated and acts as a tumor suppressor in BRAF -mutated colorectal cancer

doi: 10.1186/s12885-017-3739-x

Figure Lengend Snippet: miR-193a-3p functions as a tumor-suppressor in colorectal cancer cells and its expression was decreased by overexpression of a BRAF protein. a Transfection of precursors of miR-193a-3p increased the expression of mature miR-193a-3p (top), and inhibited cell viability of the three cell lines irrespective of their KRAS/BRAF mutational status (bottom). b, c Transfection of the precursors of miR-193a-3p inhibited cell invasion ability in RKO and HCT116 cells. d miR-193a-3p overexpression reduced the mRNA levels of EMT-related genes ZEB1 , SNAI1, and SNAI2 in RKO cells. e The mutant BRAF (V600E) overexpression activates its downstream pathway. Western blot analysis shows that the overexpression of mutant BRAF protein caused an increase in phosphorylated levels of BRAF and its downstream MEK and ERK, in the HCT8, LIM2405, SW48 and DiFi colorectal cancer cells compared to those transfected with a control vector. f miR-193a-3p expression was decreased in the KRAS/BRAF -wild-type SW48 and DiFi cells, but not in the KRAS -mutant HCT8 cells and the BRAF -mutant LIM2405 cells 72 h after overexpression of mutant BRAF protein. The Student’s t test was used to analyze statistical differences

Article Snippet: Anti-BRAF rabbit monoclonal antibody (#9433, Cell Signaling Technology, Danvers, MA, USA), anti-p-BRAF (Ser445) rabbit monoclonal antibody (#2696, Cell Signaling Technology), anti-p-MEK1/2 (Ser217/221) rabbit polyclonal antibody (#9121, Cell Signaling Technology), anti-p-ERK1/2 (Ser217/221) rabbit monoclonal antibody (#4094, Cell Signaling Technology), and anti-α-tubulin mouse monoclonal antibody (Sigma-Aldrich) were used as primary antibodies for detection of the specific proteins.

Techniques: Expressing, Over Expression, Transfection, Mutagenesis, Western Blot, Control, Plasmid Preparation

Journal: BMC Cancer

Article Title: microRNA-193a-3p is specifically down-regulated and acts as a tumor suppressor in BRAF -mutated colorectal cancer

doi: 10.1186/s12885-017-3739-x

Figure Lengend Snippet: Low miR-193a-3p expression is associated with a worse survival of colorectal cancer patients treated with anti-EGFR therapy. Kaplan–Meyer curves for a OS from the start of anti-EGFR therapy and b PFS based upon the KRAS/BRAF mutational status ( n = 45). Kaplan-Meyer curves for c OS and d PFS based upon miR-193a-3p expression in patients with any KRAS/BRAF mutational status (n = 45). Kaplan-Meyer curves for e OS and f PFS based upon miR-193a-3p expression in the KRAS/BRAF -wild-type group ( n = 34)

Article Snippet: Anti-BRAF rabbit monoclonal antibody (#9433, Cell Signaling Technology, Danvers, MA, USA), anti-p-BRAF (Ser445) rabbit monoclonal antibody (#2696, Cell Signaling Technology), anti-p-MEK1/2 (Ser217/221) rabbit polyclonal antibody (#9121, Cell Signaling Technology), anti-p-ERK1/2 (Ser217/221) rabbit monoclonal antibody (#4094, Cell Signaling Technology), and anti-α-tubulin mouse monoclonal antibody (Sigma-Aldrich) were used as primary antibodies for detection of the specific proteins.

Techniques: Expressing

Journal: BMC Cancer

Article Title: microRNA-193a-3p is specifically down-regulated and acts as a tumor suppressor in BRAF -mutated colorectal cancer

doi: 10.1186/s12885-017-3739-x

Figure Lengend Snippet: Up-regulated and down-regulated miRNAs of BRAF -mutant colorectal cancer samples screened using a miRNA microarray analysis

Article Snippet: Anti-BRAF rabbit monoclonal antibody (#9433, Cell Signaling Technology, Danvers, MA, USA), anti-p-BRAF (Ser445) rabbit monoclonal antibody (#2696, Cell Signaling Technology), anti-p-MEK1/2 (Ser217/221) rabbit polyclonal antibody (#9121, Cell Signaling Technology), anti-p-ERK1/2 (Ser217/221) rabbit monoclonal antibody (#4094, Cell Signaling Technology), and anti-α-tubulin mouse monoclonal antibody (Sigma-Aldrich) were used as primary antibodies for detection of the specific proteins.

Techniques: Mutagenesis, Microarray