Journal: Cell Reports

Article Title: Aryl Hydrocarbon Receptor Contributes to the Transcriptional Program of IL-10-Producing Regulatory B Cells

doi: 10.1016/j.celrep.2019.10.018

Figure Lengend Snippet:

Article Snippet: Negative CD43- Isolation Kit , Miltenyi Biotec , Cat# 130-049-801.

Techniques: Western Blot, Recombinant, Methylation, Adjuvant, Enzyme-linked Immunosorbent Assay, Isolation, cDNA Synthesis, SYBR Green Assay, DNA Library Preparation, Purification, Bicinchoninic Acid Protein Assay, Microarray, Ex Vivo, Generated, Software, Red Blood Cell Lysis, Staining, Lysis

Journal: Molecular Medicine Reports

Article Title: Alterations in the long non-coding RNA transcriptome in mesangial cells treated with aldosterone in vitro

doi: 10.3892/mmr.2017.7313

Figure Lengend Snippet: Reverse transcription-quantitative polymerase chain reaction quantification of microarray hybridization. (A) The relative expression level of lncRNAs BC168211, BC088254, AF336872, AY325162, BC168687, AF230638 and BC167085. (B) The relative expression levels of upregulated mRNAs, NM_001108598, NM_001109190 and NM_001101018, and (C) downregulated mRNAs, NM_019347, NM_177,962 and NM_0,011,08823. *P<0.05 vs. the control group. ALD, aldosterone; lncRNAs, long non-coding RNAs; MCs, mesangial cells.

Article Snippet: The cells were stored at −80°C and sent to the LncRNA Expression Microarray (Arraystar, Rockville, MD, USA).

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, Microarray, Hybridization, Expressing, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: M 6 A -mediated lncRNA SCIRT stability promotes NSCLC progression through binding to SFPQ and activating the PI3K/Akt pathway

doi: 10.1007/s00018-025-05594-z

Figure Lengend Snippet: SCIRT m 6 A modification and expression levels were elevated in NSCLC. ( a ) Heatmap of differentially m 6 A-methylated lncRNAs. ( b, c ) Differentially m 6 A-methylated lncRNA-related mRNAs were analyzed by GO and the KEGG using TCGA data. ( d ) SCIRT, RP11-385J1.2, and SNHG9 m 6 A methylation levels in 10 paired NSCLC and adjacent noncancerous tissues. ( e ) Relative expression levels of SCIRT in 10 paired NSCLC and adjacent noncancerous tissues. ( f ) SCIRT expression levels of NSCLC (A549, H1299, and H358) and bronchial epithelial cell lines (BEAS-2B). Data are presented as mean ± SD from at least three independent experiments. ** P < 0.01, *** P < 0.001

Article Snippet: Arraystar m 6 A lncRNA epitranscriptomic microarrays were used to screen differentially m 6 A-methylated lncRNAs in NSCLC, and array figures were shown in Supplementary Fig. . A total of 39 differential metylated candidates were identified with 24 upregulated genes and 15 downregulated genes (Table ).

Techniques: Modification, Expressing, Methylation

Journal: Nucleic Acids Research

Article Title: A transcriptomic and proteomic map of primary human cell types

doi: 10.1093/nar/gkaf1498

Figure Lengend Snippet: Cell type specific genes demonstrated at mRNA level. (A) Heat map of genes with cell type specific expression. (B) Heat map of selected genes with endothelial or epithelial cell type specific expression. (C) Immunohistochemistry staining of protein C1orf116 across human tissues showing its epithelial cell type specific expression. The panel shows a magnified view (20×) from tissue microarrays.

Article Snippet: The Human Protein Atlas (HPA) project provided a tissue-based map of the human proteome through transcriptomics and tissue microarray-based immunohistochemistry analysis [ , ].

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Oncogene

Article Title: Effective targeting of breast cancer stem cells by combined inhibition of Sam68 and Rad51

doi: 10.1038/s41388-022-02239-4

Figure Lengend Snippet: A Workflow showing the selection strategy for KHDRBS1 among DNA-damage response genes transcriptionally activated by Myc and significantly associated to breast cancer prognosis. Venn diagram showing the overlap between Myc-transcriptionally activated genes, DNA-damage response genes and genes associated to breast cancer. Specifically, genes were retrieved from: (i) microarray data of Myc-overexpressing mammospheres (M2) (GSE86407); (ii) published dataset (MD Anderson Human-DNA Repair Genes, https://www.mdanderson.org/documents/Labs/Wood-Laboratory/human-dna-repair-genes.html ), BioRad DNA-damage signaling pathway (SAB Target List H96) and recently published DNA-damage-associated genes (Supplementary Table ); and (iii) breast cancer versus normal breast tissues TCGA BRCA and GTeX gene expression data (Supplementary Table ). Genes were further selected for association to the worse relapse-free survival probability in breast cancer (Supplementary Table ) and novelty in the field, excluding known genes associated with BRCAness . B Box plot representing the distribution of log2 gene expression of KHDRBS1 retrieved from TCGA BRCA ( n = 1212) and GTeX ( n = 179) gene expression data (RNASeq2GeneNorm). p value was calculated with Wilcoxon rank sum test. C Kaplan–Meier plots of relapse-free survival (RFS) probability of BC patients stratified by high or low KHDRBS1 expression levels. D GSEA of DNA-repair gene signatures in IMEC-WT versus M2 ( n =3). E Scheme showing MYC and H3K4me3 PCR amplicons localization (red box) on IMEC-WT and M2 cells and layered H3K27ac signals on KHDRBS1 ( SAM68 ) promoter from ENCODE. Chromatin state was assessed by ChromHMM from ENCODE. MYC-MAX binding on multiple cell lines was assessed by ChIP-seq from ENCODE. F ChIP-qPCR estimating MYC binding at SAM68 promoter in IMEC-WT and M2 cells. Data are mean ± SEM ( n = 3). G qRT-PCR analysis of SAM68 gene expression in IMEC-WT and M2 cells. Data are mean ± SEM ( n = 3). H ChIP-qPCR of H3K4me3 deposition at KHDRBS1 ( SAM68 ) promoter in IMEC-WT and M2 cells. Data are mean ± SEM ( n = 3).

Article Snippet: For immunoprecipitation experiments, an equal amount of protein lysates was incubated overnight at 4 °C with 2 μg of anti-Sam68 antibody (H-4, mouse IgG 1 , Santa Cruz Biotechnology) mixed with protein G-Sepharose (Sigma-Aldrich).

Techniques: Selection, Microarray, Gene Expression, Expressing, Binding Assay, ChIP-sequencing, ChIP-qPCR, Quantitative RT-PCR

Journal: Oncogene

Article Title: Effective targeting of breast cancer stem cells by combined inhibition of Sam68 and Rad51

doi: 10.1038/s41388-022-02239-4

Figure Lengend Snippet: A Kaplan–Meier plots of distant relapse-free survival (DRFS) of BC patients stratified by high or low Sam68 protein expression levels. Patients were categorized according to all molecular subtypes ( n = 211) and Luminal-A ( n = 91), Luminal-B ( n = 61), HER2 + ( n = 27), TNBC ( n = 32), HER2 + + TNBC ( n = 59) BCs. B Box plot representing the distribution of log2 gene expression of KHDRBS1 retrieved from TCGA BRCA gene expression data (RNASeq2GeneNorm). p value was calculated with Wilcoxon rank sum test. The indicated statistics refer to each molecular subtype versus basal subtypes. * p value ≤ 0.05; ** p value ≤ 0.01; **** p value ≤ 0.0001. C ChIP-qPCR estimating MYC and MAX binding at SAM68 promoter in BCSphCs (#4 and #15). Data are mean ± SEM of two independent experiment for each BCSphCs. D Expression of Myc (green color) and Sam68 (red color) on paraffin-embedded sections on parental BC and corresponding PDX tissue. Nuclei were counterstained with Toto-3 (blue color). Scale bar represents 40 µm. E Relative mRNA expression levels of MYC and KHDRBS1 on BCSphCs (#4, #13, and #21) expressing a MycER fusion protein induced by 50 nM of OHT. Data are represented as fold mRNA level changes of OHT-treated cells over vehicle. Data are represented as mean ± SD of three independent experiments. * p value ≤ 0.05; ** p value ≤ 0.01. F Cell proliferation analysis of ER+ (MCF7), TNBC (BT549), TNBC BRCA mut (HCC1937) BC cell lines and BCSphCs (#1, #4, #13, and #21) transduced with doxycyclin-inducible non-targeting (nt) and short hairpin Sam68 (shSam68). Data are represented as fold variation of shSam68 over scr. ns not significant; ** p value ≤ 0.01. G Size of tumors generated by orthotopic injection of ER+ (MCF7), TNBC (BT549), TNBC BRCA mut (HCC1937) BC cell lines and BCSphCs (#4, #13) in immunocompromised mice (NOD/SCID) at the indicated time points. Data are expressed as mean ± SD ( n = 5 mice per group). ns not significant, *** p value ≤ 0.001.

Article Snippet: For immunoprecipitation experiments, an equal amount of protein lysates was incubated overnight at 4 °C with 2 μg of anti-Sam68 antibody (H-4, mouse IgG 1 , Santa Cruz Biotechnology) mixed with protein G-Sepharose (Sigma-Aldrich).

Techniques: Expressing, Gene Expression, ChIP-qPCR, Binding Assay, Transduction, Generated, Injection

Journal: Oncogene

Article Title: Effective targeting of breast cancer stem cells by combined inhibition of Sam68 and Rad51

doi: 10.1038/s41388-022-02239-4

Figure Lengend Snippet: A MYC binding on DNA-damage related genes transcription start sites (TSS) on IMEC-WT and M2 breast cells. B Representative immunofluorescence analysis of Rad51 foci formation in ER+ (MCF7), TNBC (BT549), TNBC BRCA mut (HCC1937) BC established cell lines and BCSphCs (#4) untreated (UT) and after 6 h of 8 Gy single dose γ-irradiation (IR). Nuclei were counterstained by Toto-3 (blue). Scale bar represents 10 µm. C Waterfall plot analysis of doxorubicin (DOX, 200 nM, left panel ), paclitaxel (PTX, 10 nM, middle panel ) and carboplatin (CARB, 100 µM, left panel ) response at 72 h in ER+ and TNBC BC established cell lines and BCSphCs. D Response rate distribution to chemotherapy for ER+ and TNBC BC established cell lines and BCSphCs treated as in ( C ). Middle line shows the median value of response per group, while single points represent the average value of BC cell response to DOX, PTX and CARB. Data are mean of three independent experiments. Statistical analysis was performed by using Kruskal–Wallis test. Ns not significant, * p value ≤ 0.05; ** p value ≤ 0.01. E Immunoblot analysis of PARP and Sam68 (input) and after immunoprecipitation (IP) with Sam68 antibody in BCSphCs (#15) treated for 4 h with vehicle, doxorubicin (DOX), paclitaxel (PTX) and carboplatin (CARB). Lamin-B was used as loading control. F Immunoblot analysis of nuclear PAR, PARP, and Sam68 in scramble (scr) and short hairpin Sam68 (shSam68) ER+ (MCF7), TNBC (BT549), and TNBC BRCA mut (HCC1937) BC cell lines and BCSphCs (#4) treated with vehicle, doxorubicin (DOX), paclitaxel (PTX) and carboplatin (CARB) for 4 h. H3 was used as loading control. G Cell proliferation analysis of ER+ (MCF7), TNBC (BT549), and TNBC BRCA mut (HCC1937) BC cell lines and BCSphCs (#1, #4, #13, #21) transduced with scramble and short hairpin Sam68 (shSam68) treated with vehicle, doxorubicin (DOX), paclitaxel (PTX) and carboplatin (CARB) for 72 h. Data are represented as fold variation of shSam68 over scramble. Data are mean ± SD of three independent experiments. ns not significant; * p value ≤ 0.05; ** p value ≤ 0.01. H , I Relative mRNA expression levels of RAD51 (H) and MYC (I) on scramble (scr) and short hairpin Sam68 (shSam68) ER+ (MCF7), TNBC (BT549), and TNBC BRCA mut (HCC1937) BC cell lines and BCSphCs (#12 and #13) treated with vehicle, doxorubicin (DOX), paclitaxel (PTX), and carboplatin (CARB) for 24 h. Data are represented as fold mRNA level changes of treated scr and shSam68 cells over vehicle. Data are represented as mean ± SD of three independent experiments. Ns not significant, * p value ≤ 0.05; ** p value ≤ 0.01; *** p value ≤ 0.001.

Article Snippet: For immunoprecipitation experiments, an equal amount of protein lysates was incubated overnight at 4 °C with 2 μg of anti-Sam68 antibody (H-4, mouse IgG 1 , Santa Cruz Biotechnology) mixed with protein G-Sepharose (Sigma-Aldrich).

Techniques: Binding Assay, Immunofluorescence, Irradiation, Western Blot, Immunoprecipitation, Control, Transduction, Expressing

Journal: Oncogene

Article Title: Effective targeting of breast cancer stem cells by combined inhibition of Sam68 and Rad51

doi: 10.1038/s41388-022-02239-4

Figure Lengend Snippet: A Schematic model of DNA-repair signaling pathways mediating the resistance of BC stem-like cells to chemotherapy. B Workflow of purification of sphere cells from serially transplanted BC PDX and their use for in vitro and in vivo drug toxicity testing. C Size of tumors generated by orthotopic injection of scramble (scr) and short hairpin Sam68 (shSam68) BCSphCs treated with vehicle (veh) and BO2. Arrows indicate the start and the end of treatment. Data are expressed as mean of tumors generated by the injection of BCSphCs (#4, #13, and #21) ± SEM ( n = 5 mice per group). D Size of tumors generated by orthotopic injection of scramble (scr) and short hairpin Sam68 (shSam68) BCSphCs (#4, #13, #21) treated with vehicle, olaparib, BO2, cisplatin and olaparib plus BO2 and olaparib plus cisplatin and BO2. Arrows indicate the beginning and the end of treatment. Data are expressed as mean of tumors generated by the injection of BCSphCs (#4, #13, and #21) ± SEM ( n = 5 mice per group). **** p value ≤ 0.0001. E Immunoblot analysis of Rad51 in BCSphCs (#15) treated with dinaciclib for 24 h at the indicated concentration. Β-actin was used as loading control. F Cell viability percentage of scramble (scr) and short hairpin Sam68 (shSam68) BCSphCs (#4, #13, #15, and #21) treated with vehicle and dinaciclib (10 nM) for 6 days. Data are represented as mean ± SEM ( n = 2). * p value ≤ 0.05; *** p value ≤ 0.001. G Representative images ( left panel ) and quantification of area ( right panel ) of BC sphere cells (#21), transduced with scramble (scr) and short hairpin Sam68 (shSam68) lentiviral vectors, treated with vehicle and dinaciclib for 6 days. Data are represented as mean ± SEM ( n = 3). Ns not significant, ** p value ≤ 0.01; *** p value ≤ 0.001. Scale bar represents 100 µm. H Size of tumors generated by orthotopic injection of scramble (scr) and short hairpin Sam68 (shSam68) BCSphCs treated with vehicle (veh) and dinaciclib (din). Arrows indicate the start and the end of treatment. Data are expressed as mean of tumors generated by the injection of BCSphCs (#4, #7, #13) ± SEM ( n = 5 mice per group). **** p value ≤ 0.0001. I Cell viability percentage of BCSphCs (#4, #13, #14, #15, #21) treated with vehicle, olaparib and dinaciclib, alone or in combination, at the indicated concentrations for 6 days. Data are represented as mean ± SD ( n = 3). J Synergy plot representing the combination index (CI), computed in CompuSyn by using Chou-Talalay method, for each olaparib and dinaciclib dose pair, calculated from cell viability data of BCSphCs (#13). K Size of tumors generated by orthotopic injection of BCSphCs treated with vehicle, olaparib, dinaciclib and olaparib plus dinaciclib. Arrows indicate the start and the end of treatment. Data are expressed as mean of tumors generated with BCSphCs (#4, #7, #13) ± SEM ( n = 5 mice per group). *** p value ≤ 0.001.

Article Snippet: For immunoprecipitation experiments, an equal amount of protein lysates was incubated overnight at 4 °C with 2 μg of anti-Sam68 antibody (H-4, mouse IgG 1 , Santa Cruz Biotechnology) mixed with protein G-Sepharose (Sigma-Aldrich).

Techniques: Protein-Protein interactions, Purification, In Vitro, In Vivo, Generated, Injection, Western Blot, Concentration Assay, Control, Transduction

Journal: Oncogene

Article Title: Effective targeting of breast cancer stem cells by combined inhibition of Sam68 and Rad51

doi: 10.1038/s41388-022-02239-4

Figure Lengend Snippet: A Cell viability percentage of scramble (scr) and short hairpin Sam68 (shSam68) ER+ R (MCF7) BC cell line treated with vehicle and dinaciclib (10 nM) for 6 days. Data are represented as mean ± SEM ( n = 4). * p value ≤ 0.05; ** p value ≤ 0.01; *** p value ≤ 0.001. B Relative mRNA expression levels of RAD51 and MYC on scramble (scr) and short hairpin Sam68 (shSam68) ER+ R (MCF7) BC cells treated with vehicle and dinaciclib for 6 days. Data are represented as fold mRNA level changes of treated scr and shSam68 over vehicle ( n = 3). C Cell viability percentage in ER+ R (MCF7) BC cells treated with vehicle, olaparib and dinaciclib, alone or in combination, at the indicated concentrations for 6 days. Data are represented as mean ± SD ( n = 3). D Kaplan–Meier plots of relapse-free survival (RFS) probability of BC patients of all molecular subtypes stratified by high or low MYC , KHDRBS1 , and RAD51 expression levels. E Schematic model showing the persistence of a BC stem-like population, characterized by high expression levels of MYC, SAM68 , and RAD51 , following standard anticancer therapies.

Article Snippet: For immunoprecipitation experiments, an equal amount of protein lysates was incubated overnight at 4 °C with 2 μg of anti-Sam68 antibody (H-4, mouse IgG 1 , Santa Cruz Biotechnology) mixed with protein G-Sepharose (Sigma-Aldrich).

Techniques: Expressing

Journal: Molecular cell

Article Title: Harnessing BET Inhibitor Sensitivity Reveals AMIGO2 as a Melanoma Survival Gene

doi: 10.1016/j.molcel.2017.11.004

Figure Lengend Snippet: (A) Immunofluorescence of SKmel147 cells stably expressing AMIGO2-GFP (green), stained with AMIGO2 antibody (red) and Hoechst 33342 (blue). Scale bar, 20 μm. (B) Functional annotation of AMIGO2-interacting proteins detected by GFP pull-down followed by MS in SKmel147 cells stably expressing AMIGO2-GFP (see Table S4). (C) PTK7 and GFP immunoblots following GFP pull-down from 501MEL cells stably expressing AMIGO2-GFP. (D) Full-length PTK7 (FL-PTK7), C-terminal fragments CTF1- and CTF2-PTK7, and FOXM1 immunoblots of 501MEL cells 72 hr post-infection with shSCR or shPTK7 (shP7 #1 and #2). Actin was used as a loading control. (E) Relative growth curves of 501MEL (left) and SKmel147 (right) cells stably transduced with shSCR or shPTK7 (shP7 #1 and #2). Values are normalized to seeding control (n = 3). (F) Percent Annexin V-positive cells 6 days post-transduction for same cells as in (E). (G) FL-PTK7, CTF-PTK7, and FOXM1 immunoblots of 501MEL cells 48 hr post-transduction with shSCR or shAMIGO2 (shA2 #1 and #2). Actin was used as a loading control. (H) FL-PTK7, CTF-PTK7, FOXM1, and AMIGO2 immunoblots of 501MEL cells untreated or treated with JQ1 (JQ1[+]) for 72 hr. Tubulin was used as a loading control. (I) CTF2-PTK7 immunoblot of nuclear lysates from same cell as in (G) (left). Lamin B1 was used as loading control. Signal quantification (right), normalized to Lamin B1, relative to shSCR (n = 3). All values and error bars represent mean ± SD or ± SEM. See also Figures S3 and S4.

Article Snippet: LAMIN B1 , Santa Cruz , SC-6217.

Techniques: Immunofluorescence, Stable Transfection, Expressing, Staining, Functional Assay, Western Blot, Infection, Control, Transduction

Journal: Molecular cell

Article Title: Harnessing BET Inhibitor Sensitivity Reveals AMIGO2 as a Melanoma Survival Gene

doi: 10.1016/j.molcel.2017.11.004

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: LAMIN B1 , Santa Cruz , SC-6217.

Techniques: Microarray, Derivative Assay, Recombinant, Magnetic Beads, Flow Cytometry, Caspase Activity Assay, DNA Library Preparation, Blocking Assay, Extraction, TA Cloning, Sequencing, RNA Sequencing, Western Blot, Mass Spectrometry, Expressing, Software

Journal: BMC Cancer

Article Title: Novel HDGF/HIF-1α/VEGF axis in oral cancer impacts disease prognosis

doi: 10.1186/s12885-019-6229-5

Figure Lengend Snippet: Correlation of HDGF and VEGF expression in oral cancer. a , b Correlation between HDGF and VEGF mRNA levels in oral cancer and head and neck cancer patients obtained by analysis of data from TCGA. HDGF expression is positively correlated with VEGFA expression in human head and neck squamous cell carcinoma tissues, including oral cancer. c Tissue microarray analysis of the correlation between HDGF and VEGF expression in oral cancer patients. The photographs were from two representative oral cancer patients. Case 1 (pT2N0M0, stage II) exhibited low-expression HDGF and VEGF immunostaining, whereas Case 2 (pT3N2M0, stage III) showed high-expression staining of both HDGF and VEGF. Scale bars, 20 μm. d Immunofluorescence staining of oral cancer patients. The fluorescent color of HDGF was Green (AlexaFluor 488); VEGF was red (AlexaFluor 546); nuclei were stained with blue color (DAPI). Case 3 (pT1N0M0, stage I) exhibited both high-intensity staining of HDGF and VEGF, whereas Case 4 (pT2N0M0, stage II) showed intermediate-intensity HDGF and VEGF immunofluorescence staining, and Case 5 (pT1N0M0, stage II) showed low-intensity staining of HDGF and VEGF. Scale bars, 20 μm

Article Snippet: Briefly, the slides were incubated with primary HDGF antibody (1:200 dilution) and VEGF antibodies (1:250; Santa Cruz; Santa Cruz, CA, USA) for 30 min and visualized using a peroxidase-conjugated secondary antibody, a polymer detection system (Zymed Laboratories, San Francisco, CA, USA) and 3,3-diaminobenzidine tetrahydrochloride (Sigma, St. Louis, MO).

Techniques: Expressing, Microarray, Immunostaining, Staining, Immunofluorescence

Journal: BMC Cancer

Article Title: Novel HDGF/HIF-1α/VEGF axis in oral cancer impacts disease prognosis

doi: 10.1186/s12885-019-6229-5

Figure Lengend Snippet: Effect of HDGF on VEGF expression in oral cancer cells. SCC4 cells were treated with the indicated concentration of recombinant HDGF protein for 24 h before harvest. a Relative gene expression levels of VEGF were analyzed by SYBR green-based RT-PCR. Data are expressed as the fold change with respect to the control group (means ± SD of triplicate experiments). b Cell lysates were analyzed using Western blotting, and the protein levels of VEGF/β-actin were measured and quantified. c The secreted VEGF protein levels in the supernatants were measured by Western blotting. Ponceau S staining was used as a loading control. d Levels of secreted VEGF protein (pg/ml) were detected by enzyme-linked immunosorbent assay (ELISA) in triplicate experiments. Data were mean of three experiments. *, P < 0.05; **, P < 0.01; ns, not statistically significant

Article Snippet: Briefly, the slides were incubated with primary HDGF antibody (1:200 dilution) and VEGF antibodies (1:250; Santa Cruz; Santa Cruz, CA, USA) for 30 min and visualized using a peroxidase-conjugated secondary antibody, a polymer detection system (Zymed Laboratories, San Francisco, CA, USA) and 3,3-diaminobenzidine tetrahydrochloride (Sigma, St. Louis, MO).

Techniques: Expressing, Concentration Assay, Recombinant, Gene Expression, SYBR Green Assay, Reverse Transcription Polymerase Chain Reaction, Control, Western Blot, Staining, Enzyme-linked Immunosorbent Assay

Journal: BMC Cancer

Article Title: Novel HDGF/HIF-1α/VEGF axis in oral cancer impacts disease prognosis

doi: 10.1186/s12885-019-6229-5

Figure Lengend Snippet: The Neutralizing antibody against nucleolin eliminate HDGF-stimulated AKT/HIF-1α/NF-κB/VEGF signaling in SCC4 oral cancer cells. a - d SCC4 cells were treated with recombinant HDGF protein (100 ng/ml) in the presence of anti-NCL or anti-IgG antibody (5 μg/ml) for 24 h before total protein extraction. Cell lysates were subjected to Western blotting with the indicated antibodies. β-actin was used as an internal control for loading and transfer. Data were mean of three experiments. *, P < 0.05; **, P < 0.01; ns, not statistically significant

Article Snippet: Briefly, the slides were incubated with primary HDGF antibody (1:200 dilution) and VEGF antibodies (1:250; Santa Cruz; Santa Cruz, CA, USA) for 30 min and visualized using a peroxidase-conjugated secondary antibody, a polymer detection system (Zymed Laboratories, San Francisco, CA, USA) and 3,3-diaminobenzidine tetrahydrochloride (Sigma, St. Louis, MO).

Techniques: Recombinant, Protein Extraction, Western Blot, Control

Journal: BMC Cancer

Article Title: Novel HDGF/HIF-1α/VEGF axis in oral cancer impacts disease prognosis

doi: 10.1186/s12885-019-6229-5

Figure Lengend Snippet: Effects of chetomin and Bay 11–7082 on HDGF-induced VEGF upregulation in SCC4 oral cancer cells. Cells were treated with recombinant HDGF protein (100 ng/ml) in the presence of Bay 11–7082 (10 nM) or chetomin (10 nM) for 24 h. a Relative gene expression levels of VEGF were analyzed by SYBR Green-based RT-PCR. Data are expressed as the fold change with respect to the control group (means ± SD of triplicate experiments). b The protein levels of VEGF were analyzed by Western blotting and normalized to β-actin expression. ( c ) the levels of secreted VEGF protein (pg/ml) were detected by ELISA in triplicate experiments. d Scheme for HDGF-regulated VEGF transcription in oral cancer cells. Data were mean of three experiments. *, P < 0.05; **, P < 0.01; ns, not statistically significant

Article Snippet: Briefly, the slides were incubated with primary HDGF antibody (1:200 dilution) and VEGF antibodies (1:250; Santa Cruz; Santa Cruz, CA, USA) for 30 min and visualized using a peroxidase-conjugated secondary antibody, a polymer detection system (Zymed Laboratories, San Francisco, CA, USA) and 3,3-diaminobenzidine tetrahydrochloride (Sigma, St. Louis, MO).

Techniques: Recombinant, Gene Expression, SYBR Green Assay, Reverse Transcription Polymerase Chain Reaction, Control, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay

Journal: BMC Cancer

Article Title: Novel HDGF/HIF-1α/VEGF axis in oral cancer impacts disease prognosis

doi: 10.1186/s12885-019-6229-5

Figure Lengend Snippet: Multivariate analyses of HDGF and VEGF

Article Snippet: Briefly, the slides were incubated with primary HDGF antibody (1:200 dilution) and VEGF antibodies (1:250; Santa Cruz; Santa Cruz, CA, USA) for 30 min and visualized using a peroxidase-conjugated secondary antibody, a polymer detection system (Zymed Laboratories, San Francisco, CA, USA) and 3,3-diaminobenzidine tetrahydrochloride (Sigma, St. Louis, MO).

Techniques: Expressing

Journal: Cell Reports Medicine

Article Title: Discovery of galectin-8 as an LILRB4 ligand driving M-MDSCs defines a class of antibodies to fight solid tumors

doi: 10.1016/j.xcrm.2023.101374

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-HA Tag (Clone F-7) , Santa Cruz Biotechnology , Cat#sc-7392; RRID: AB_627809.

Techniques: Recombinant, Derivative Assay, Microarray, Enzyme-linked Immunosorbent Assay, Lysis, Blocking Assay, Staining, Transfection, Marker, Flow Cytometry, shRNA, Plasmid Preparation, Negative Control, Silver Staining, Bicinchoninic Acid Protein Assay, Selection, Extraction, Labeling, RNA Expression, Cell Culture, Software