il 6 duoset elisa kits  (R&D Systems)

Journal: The Journal of Biological Chemistry

Article Title: Novel markers of MCL1 inhibitor sensitivity in triple-negative breast cancer cells

doi: 10.1016/j.jbc.2024.107375

Figure Lengend Snippet: A 4 gene signature (GS) correlates with AZD5991 sensitivity and MCL-1 dependency in TNBC cell lines. A , analysis of gene expression correlation with TIMER software in 139 TNBC tumors (TCGA) showed that the 4 genes positively correlate with each other. B , 18 TNBC cell lines from GDSC are divided into AZD5991 resistant ( green ) and sensitive ( yellow ) groups. mRNA levels (RPKM) of AXL, ETS1, IL6, and EFEMP1 in all cell lines were extracted from CCLE RNAseq database. The median level of each gene is calculated and listed on top. Gene levels above the median is marked in red and scored 1 and below median is scored 0. The sum of the 4 gene scores is listed on right . C , correlation of GS scores with AZD5991 sensitivity was analyzed with ROC curve with sensitive cells defined as event 1. The results showed a significant correlation ( p = 0.002) with an ROC area 0.95 (ROC area 1.0 indicates perfect correlation). D, 13 TNBC cell lines with MCL-1 CRISPR (DepMap public 23Q2) are divided into MCL-1 dependent ( green ) and independent ( yellow ) groups based on their Chromos scores. Gene scores are calculated as described in B. Correlation of GS scores with MCL-1 dependency was analyzed with ROC curve ( E ). The results showed a significant correlation ( p = 0.04) with a ROC area 0.79).

Article Snippet: Recombinant EFEMP1 and IL6 proteins are obtained from R&D systems.

Techniques: Expressing, Software, CRISPR

Journal: The Journal of Biological Chemistry

Article Title: Novel markers of MCL1 inhibitor sensitivity in triple-negative breast cancer cells

doi: 10.1016/j.jbc.2024.107375

Figure Lengend Snippet: Inhibition of ETS1 reduces AXL, IL6, and EFEMP1 expression and increases MCL1 inhibitor sensitivity via expression of BCL-2. A , HS578T and MDA231 cells were transfected with control siRNA or ETS1 siRNA for 24 h. B, HS578T and MDA231 cells were treated with vehicle or TK216 (1 μM) for 24 h. mRNA were qPCR analyzed for the indicated genes. Average (triplicate) relative mRNA is presented with SD indicated. Asterisks indicate statistical significance. Representative results of two independent experiments. C , HS578T and MDA231 cells were treated with vehicle, TK216 (1μM), S63845 (1μM), or TK216 + S63845 for 3 days and then analyzed for sub-G1 cells. % sub-G1 cells are presented. Asterisks indicate statistical significance. Representative results of three independent experiments. D , MDA231 cells were treated with vehicle, S63845 (1μM), or S63845 plus BGB324 (1 μM) or TK216 (1μM), for 24 h. Lysates were immunoblotted for the indicated proteins. Representative results of two independent experiments. Original images are presented in Fig. S5 . E , The cells were treated with vehicle or the ERK inhibitor ulixertinib (2 μM) for 24 h. Lysates were immunoblotted for the indicated proteins. Representative results of two independent experiments. Original images are presented in Fig. S6 . F , MDA231 cells were treated with vehicle, S63845 (1 μM), or S63845 plus ABT-199 (2 μM and 5 μM) for 72 h. G , The cells were transfected with control siRNA or the indicated siRNAs against AXL, ETS1, IL6, or EFEMP1 and then treated with vehicle or S63845 for 72 h. Cells were analyzed with FACS for the cell cycle. The average % Sub-G1 cells were presented with SD indicated. Asterisks indicate statistical significance. Representative results of two independent experiments.

Article Snippet: Recombinant EFEMP1 and IL6 proteins are obtained from R&D systems.

Techniques: Inhibition, Expressing, Transfection, Control

Journal: The Journal of Biological Chemistry

Article Title: Novel markers of MCL1 inhibitor sensitivity in triple-negative breast cancer cells

doi: 10.1016/j.jbc.2024.107375

Figure Lengend Snippet: Overexpression of ETS1 and exogenous EFEMP1 and IL6 increases BCL2 expression and promotes MCL1 inhibitor resistance. A , control BT20 cells and BT20-pLIX-ETS1 cells were treated with vehicle or doxycycline (DOX, 0.5 μg/ml) for 24 h. Lysates were immunoblotted for the indicated proteins. Representative images of two independent experiments. B , control BT20 cells and BT20-pLIX-ETS1 cells were treated with vehicle or doxycycline (DOX, 0.5 μg/ml) in combination with vehicle or S63845 for 72 h. Cells were analyzed with FACS for cell cycle. Average % Sub-G1 cells (apoptosis) were presented with SD indicated. Asterisks indicate statistical significance. Representative results of two independent experiments. BT20 cells were FBS-starved for 24 h and then treated with vehicle or recombinant EFEMP1 (50 ng/ml) for 30 min ( C ) or recombinant IL6 (50 ng/ml) for 10 min ( D ). E , BT20 cells were treated with vehicle or S63845 (1 μM) or S63845 in combination with EFEMP1 plus IL6 for 24 h. Lysates were immunoblotted for the indicated proteins. Representative results of two independent experiments. F , BT20 cells were treated with vehicle or S63845 (1 μM) or S63845 in combination with EFEMP1 plus IL6 for 72 h. Cells were analyzed with FACS for the cell cycle. Average % Sub-G1 cells (apoptosis) were presented with SD indicated. Asterisks indicate statistical significance. Representative results of two independent experiments.

Article Snippet: Recombinant EFEMP1 and IL6 proteins are obtained from R&D systems.

Techniques: Over Expression, Expressing, Control, Recombinant

Journal: iScience

Article Title: The IL-6 signaling pathway contributes critically to the immunomodulatory mechanism of human decidua-derived mesenchymal stromal cells

doi: 10.1016/j.isci.2024.109783

Figure Lengend Snippet:

Article Snippet: The test samples and standard recombinant IL-6 (R&D Systems, Minnesota, USA) were added to separate wells of the 96-well plate, followed by incubation at room temperature for 2 h. The plate was washed, biotinylated IL-6 polyclonal antibody (R&D Systems) added, and the reaction proceeded for 2 h at room temperature.

Techniques: Virus, shRNA, Plasmid Preparation, Recombinant, SYBR Green Assay, Enzyme-linked Immunosorbent Assay