Journal: Journal of Cancer

Article Title: Modulating cancer stemness provides luminal a breast cancer cells with HER2 positive-like features

doi: 10.7150/jca.37117

Figure Lengend Snippet: Expression of cell stemness and subtyping markers in stable OKMS cells. A) Expression of cell stemness markers (OCT4, KLF4, MYC and SOX2) at the A) transcriptional and B) translational levels. Expression of subtyping markers (ER, HER2) at the D) transcriptional and E) translational levels. Western blot signalling intensities normalized by that of GAPDH in OKMS cells are plotted in C) for stemness markers (OCT4, KLF4, MYC and SOX2) and in F) for subtyping markers (ER, HER2). Bars represent mean ± SD of fold change from at least three independent experiments, and ** (0.01

Article Snippet: The secondary antibody and primary antibodies against GAPDH, OCT4, KLF4, MYC, SOX2, ER, HER2 were purchased from Proteintech, and primary antibodies targeting p65, phospho-p65, ERK1/2, phospho-ERK1/2, JNK, phospho-JNK, p38, phospho-p38 MAPK were ordered from Cell Signaling Technology.

Techniques: Expressing, Western Blot

Journal: Journal of Cancer

Article Title: Modulating cancer stemness provides luminal a breast cancer cells with HER2 positive-like features

doi: 10.7150/jca.37117

Figure Lengend Snippet: Drug dose curves of cells in response to Tamoxifen and Trastuzumab. Drug response curves under the treatment of A) Tamoxifen, B) Trastuzumab. MCF7, SKBR3, MDAMB231 are luminal, HER2 positive and triple negative cells, respectively, and OKMS is a stable cell line established from MCF7 by over-expressing the four-gene panel controlling cell stemness (OCT4, KLF4, MYC and SOX2).

Article Snippet: The secondary antibody and primary antibodies against GAPDH, OCT4, KLF4, MYC, SOX2, ER, HER2 were purchased from Proteintech, and primary antibodies targeting p65, phospho-p65, ERK1/2, phospho-ERK1/2, JNK, phospho-JNK, p38, phospho-p38 MAPK were ordered from Cell Signaling Technology.

Techniques: Stable Transfection, Expressing

Journal: Oncotarget

Article Title: A function-blocking CD47 antibody suppresses stem cell and EGF signaling in triple-negative breast cancer

doi: 10.18632/oncotarget.7100

Figure Lengend Snippet: EGF A . and EGFR mRNA expression B . is up-regulated in bCSCs, and down-regulated by B6H12 antibody treatment. EGF and EGFR mRNA expressions were undetectable in bCSCs treated with B6H12 Ab. C . EGFR mRNA expression in MDA-MB-468 cells treated with B6H12 or isotype control IgG. D . Differentiated MDA-MB-231 cells and bCSCs cells were treated with B6H12 Ab for 24 h. Total miRNA was extracted, and miR-7 was analyzed using real time PCR. Relative expression of hsa-miR 7-5P was measured from differentiated and bCSCs, and SNO47 was used as control for normalization.

Article Snippet: Antibodies specific for EGFR and phospho-Tyr1175 EGFR, actin, NANOG, OCT4, and SOX2 were obtained from Cell Signaling (Danvers, MA), CD47 antibody B6H12 from (Abcam, Cambridge, MA) and Human anti-human CD47-FITC (BD Biosciences).

Techniques: Expressing, Control, Real-time Polymerase Chain Reaction

Journal: Oncotarget

Article Title: A function-blocking CD47 antibody suppresses stem cell and EGF signaling in triple-negative breast cancer

doi: 10.18632/oncotarget.7100

Figure Lengend Snippet: A. AEGFR-immunoprecipitation from MDA-MB-231 cell extracts followed by western blotting shows that B6H12 treatment for 15 min disrupts the association between EGFR and CD47 and inhibits EGFR-Y 1068 phosphorylation. B . CD47-immunoprecipitation showed that a fraction of EGFR co-immunoprecipitates with EGFR. B6H12 treatment for 15 min reduced interaction between CD47 and EGFR in MDA-MB-231 cells. ( C .- D .) MDA-MB-231 cells were pretreated with B6H12 for 15 minutes followed by EGF for 5 minutes, and IP-western blotting was performed using phospho-EGFR antibody. D . Quantification of three experiments was analyzed using the t-test (*P<0.05). E . Extracts from differentiated MDA-MB-231 cells and bCSCs treated as indicated were subjected to EGFR immunoprecipitation as in panel A. One representative blot of 3 independent experiments is shown. Numbers above lanes indicate quantiation by densitometry.

Article Snippet: Antibodies specific for EGFR and phospho-Tyr1175 EGFR, actin, NANOG, OCT4, and SOX2 were obtained from Cell Signaling (Danvers, MA), CD47 antibody B6H12 from (Abcam, Cambridge, MA) and Human anti-human CD47-FITC (BD Biosciences).

Techniques: Immunoprecipitation, Western Blot, Phospho-proteomics

Journal: International journal of oncology

Article Title: Downregulation of mitochondrial UQCRB inhibits cancer stem cell-like properties in glioblastoma.

doi: 10.3892/ijo.2017.4191

Figure Lengend Snippet: Figure 6. The effect of UQCRB inhibitors on the c‑Met signaling, HIF activity and stemness markers in glioblastoma stem‑like cells. U87MG and U373MG GSCs were treated with UQCRB inhibitors for 48 h, and the protein levels were detected by western blot analysis using specific antibodies. The levels of β-actin were used as an internal control.

Article Snippet: Anti-Nanog, anti-Sox2, anti-Oct4, anti-phospho-Met, anti-Met, anti-phospho-Stat3, anti-Stat3, anti-phospho-Akt, anti-Akt, anti-phospho-Erk1/2, anti-Erk1/2, anti-VEGF and anti-β-actin antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Activity Assay, Western Blot, Control

Journal: International journal of oncology

Article Title: Downregulation of mitochondrial UQCRB inhibits cancer stem cell-like properties in glioblastoma.

doi: 10.3892/ijo.2017.4191

Figure Lengend Snippet: Figure 10. The effect of UQCRB knockdown on the c‑Met signaling, HIF activity and stemness markers in U87MG GSCs. The protein levels were detected by western blot analysis using specific antibodies. The levels of β-actin were used as an internal control.

Article Snippet: Anti-Nanog, anti-Sox2, anti-Oct4, anti-phospho-Met, anti-Met, anti-phospho-Stat3, anti-Stat3, anti-phospho-Akt, anti-Akt, anti-phospho-Erk1/2, anti-Erk1/2, anti-VEGF and anti-β-actin antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Knockdown, Activity Assay, Western Blot, Control