Journal: BioMed Research International

Article Title: TNF α Mediated IL-6 Secretion Is Regulated by JAK/STAT Pathway but Not by MEK Phosphorylation and AKT Phosphorylation in U266 Multiple Myeloma Cells

doi: 10.1155/2013/580135

Figure Lengend Snippet: Regulation of IL-6 release by TNF α in vitro. After serum starvation, U266 and IM9 multiple myeloma cells were treated with or without 1 ng/mL TNF α for indicated times. Cell supernatants from each experimental sample were harvested for ELISA and IL-6 concentrations were determined at different time points with TNF α treatment. Relative fold changes compared to TNF α nontreated samples as a control were shown in graph. Bars represent the mean ± SEM from three independent experiments.

Article Snippet: IM9 was obtained from the Korean Cell Line Bank (Chongro-gu, Seoul, Republic of Korea).

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Control

Journal: BioMed Research International

Article Title: TNF α Mediated IL-6 Secretion Is Regulated by JAK/STAT Pathway but Not by MEK Phosphorylation and AKT Phosphorylation in U266 Multiple Myeloma Cells

doi: 10.1155/2013/580135

Figure Lengend Snippet: Effect of TNF α on activation of MAPK, JNK, and PI3K/AKT signaling pathways and effect of indicated inhibitors on IL-6 secretion. (a) U266 and IM9, multiple myeloma cell lines were starved for 8 h and then treated with or without 1 ng/mL TNF α for 10 min. Each cell line was harvested and prepared for immunoblotting. Upregulation of MAPK, JNK, and PI3K/AKT by TNF α was examined with indicated antibodies. GAPDH was used to ensure equal loading. (b) U266 cells were pretreated with various signal inhibitors for 1 h, respectively. These cells were then stimulated with TNF α for 10 min, and cell supernatants were harvested for ELISA to determine IL-6 concentration induced by TNF α . Bars represent the mean ± SEM from three independent experiments. c: control; 1: TNF α only; 2: TNF α plus 10 μ M PD98059; 3: TNF α plus 20 μ M LY294002; 4: TNF α plus 10 μ M SB203580; 5: TNF α plus 40 nM JNK inhibitor II.

Article Snippet: IM9 was obtained from the Korean Cell Line Bank (Chongro-gu, Seoul, Republic of Korea).

Techniques: Activation Assay, Protein-Protein interactions, Western Blot, Enzyme-linked Immunosorbent Assay, Concentration Assay, Control

Journal: Nature Communications

Article Title: Spatial epitranscriptomics reveals A-to-I editome specific to cancer stem cell microniches

doi: 10.1038/s41467-022-30299-3

Figure Lengend Snippet: Source data are provided as a Source Data file. a Experimental design and an example of single-cell isolation. b Single-cell isolation of the SLACS device. Scale bar, 100 µm. c Fragments per kilobase of transcript per million mapped reads (FPKM) values for the paraformaldehyde (PFA)- and methanol (MeOH)-fixed cells ( n = 60 biologically independent cells examined over 2 independent experiments). Interquartile range (IQR) of boxplot is between Q1 and Q3 and centre line indicates median value. Whiskers of boxplot is extended to the maxima and minima. Maxima is Q3 + 1.5*IQR and minima is Q1 − 1.5*IQR. d Correlation between the mRNA sequencing profiles from bulk mRNA-seq and Select-seq with ten types of PFA-fixed cells and Select-seq with ten types of MeOH-fixed cells. e Number of genes detected (FPKM) (left) and exon alignment percentage in three different cell lines fixed with PFA (right) ( n = 92 biologically independent cells examined over 3 independent experiments). Interquartile range (IQR) of boxplot is between Q1 and Q3 and centre line indicates median value. Whiskers of boxplot is extended to the maxima and minima. Maxima is Q3 + 1.5*IQR and minima is Q1 − 1.5*IQR. f Representative 3’ end bias of the full-length transcriptomes. g Principal component analysis (PCA) and h unsupervised clustering heatmap of the cells analyzed with Select-seq. i Representative transcript isoform diversity from two samples. j B cell receptor (BCR) analysis of the IM-9 cell line.

Article Snippet: Human HEK 293 T (cat # 21573), human IM-9 cells (cat # 10159), human HuT-78 cells (cat # 90078), and murine NIH3T3 cells (cat # 21658) were purchased from Korean Cell Line Bank (KCLB) and propagated according to the manufacturer’s instructions.

Techniques: Single-cell Isolation, Sequencing