Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: The STAT3-IL10-IL6 pathway is a novel regulator of macrophage efferocytosis and phenotypic conversion in sterile liver injury

doi: 10.4049/jimmunol.1701247

Figure Lengend Snippet: A. BMDMs fed with CMTMR-labelled apoT in the presence of vehicle (DMSO) or pSTAT3 (Tyr506) inhibitory peptide at various time points after adding apoT. n=3, average ± SD is reported. Data have been analyzed with two-way ANOVA for unpaired data followed by Bonferroni post-hoc test to compare all pair of columns. Data have been checked for equal variance before further analysis. *p<0.05; **p<0.01.

Article Snippet: A pSTAT3 (Tyr705) inhibitory peptide or an irrelevant peptide (MERK Millipore) were used at a 30 μM final concentration in vitro . rmIL10 was used at a final concentration of 10ng/mL(Miltenyi Biotec) ( 25 ).

Techniques:

Journal: Cancer Cell International

Article Title: Silencing of AURKA augments the antitumor efficacy of the AURKA inhibitor MLN8237 on neuroblastoma cells

doi: 10.1186/s12935-019-1072-y

Figure Lengend Snippet: MLN8237 treatment resulted in abnormal activation of Akt/Stat3 pathway. IMR32 cells were treated with 2 μmol/l of MLN8237. DMSO or no treatment as control. Cell samples were collected at 6 h and 24 h. Total proteins were extracted for western blot analysis for a Akt, pAkt, and mTOR; b GSK3β and pGSK3β; c Stat3 and pStat3. Gray values were calculated by imageJ software. The data were shown as the mean ± SEM of three independent experiments. *P < 0.05

Article Snippet: The antibodies used were as follows: anti-AURKA (abcam; ab52973; 1:30,000), anti-pAURKA(pT288) (abcam; ab52973; 1:1000), anti-MYCN (Novus Biologicals; NB200-109; 1:800), anti-pMYCN (S62) (abcam; ab185656; 1:1000), anti-MYCN (abcam; ab185655; 1:1000), anti-cyclinB (Arigo; ARG55257; 1:1000), anti-GSK3β(abcam; ab3239; 1:1000), anti-pGSK3β(Y216) (abcam; ab75745; 1:1000), anti-PTEN (abcam; ab32199; 1:10,000), anti-P27 (abcam; ab32034; 1:1000), anti-P53 (Genetex; GTX102965; 1:1000), anti-P21 (abcam; ab80633; 1:1000), anti-AKT (Arigo; A54929; 1:1000), anti-AKT(Ser473) (CST; #9271; 1:1000), anti-PI3K (abcam; ab86714; 1:1000), anti-RB (Arigo; ARG51103; 1:1000), anti-pRB (Ser795) (Arigo; ARG51631; 1:1000), anti-pRB(Ser807) (Arigo; ARG51632; 1:1000), anti-P16 (Genetex; GTX129903; 1:1000), anti-STAT3 (Genetex; GTX104616; 1:1000), anti-pSTAT3 (Tyr705) (Arigo; ARG51549; 1:1000), anti-JAK2 (Genetex; GTX101132; 1:1000), anti-pJAK2 (Y1007 + Y1008) (abcam; ab32101; 1:5000), anti-Bmi1 (Arigo; ARG55885; 1:1000), anti-survivin (Proteintech; 10508-1-AP; 1:1000), anti-GAPDH (abcam; ab8245; 1:10,000).

Techniques: Activation Assay, Western Blot, Software

Journal: Cancer Cell International

Article Title: Silencing of AURKA augments the antitumor efficacy of the AURKA inhibitor MLN8237 on neuroblastoma cells

doi: 10.1186/s12935-019-1072-y

Figure Lengend Snippet: MLN8237 treatment followed by knockdown of AURKA forced senescent cells into apoptosis. a IMR32 cells were treated with 2 μmol/l of MLN8237. Two hours later, MLN8237 withdrawal followed by siAURKA-1 transfection. At 24-h after transfection, removing the supernatant, cells were cultured in normal medium in the presence of 2 μmol/l of MLN8237 till 72 h. Cellular senescence was evaluated by SA-β-gal staining, cell apoptosis ( b ) was assessed by flow cytometry. Cell viability ( c ) was assayed by MTT method from day 1 to day 6 after 2 μmol/l of MLN8237 treatment, or siAURKA transfection alone, or MLN8237 treatment plus siAURKA transfection, or no treatment as control. Each sample was analyzed by triplicates. Error bars correspond to the averages ± S.D. d IMR32 cells were treated with 2 μmol/l of MLN8237. At 48 h after transfection, cells were harvested and total protein were isolated. Western blots were assayed for pAkt, pSTAT3, and Bmi1. Gray values were calculated by imageJ software. The data were shown as the mean ± SEM of three independent experiments. *P < 0.05

Article Snippet: The antibodies used were as follows: anti-AURKA (abcam; ab52973; 1:30,000), anti-pAURKA(pT288) (abcam; ab52973; 1:1000), anti-MYCN (Novus Biologicals; NB200-109; 1:800), anti-pMYCN (S62) (abcam; ab185656; 1:1000), anti-MYCN (abcam; ab185655; 1:1000), anti-cyclinB (Arigo; ARG55257; 1:1000), anti-GSK3β(abcam; ab3239; 1:1000), anti-pGSK3β(Y216) (abcam; ab75745; 1:1000), anti-PTEN (abcam; ab32199; 1:10,000), anti-P27 (abcam; ab32034; 1:1000), anti-P53 (Genetex; GTX102965; 1:1000), anti-P21 (abcam; ab80633; 1:1000), anti-AKT (Arigo; A54929; 1:1000), anti-AKT(Ser473) (CST; #9271; 1:1000), anti-PI3K (abcam; ab86714; 1:1000), anti-RB (Arigo; ARG51103; 1:1000), anti-pRB (Ser795) (Arigo; ARG51631; 1:1000), anti-pRB(Ser807) (Arigo; ARG51632; 1:1000), anti-P16 (Genetex; GTX129903; 1:1000), anti-STAT3 (Genetex; GTX104616; 1:1000), anti-pSTAT3 (Tyr705) (Arigo; ARG51549; 1:1000), anti-JAK2 (Genetex; GTX101132; 1:1000), anti-pJAK2 (Y1007 + Y1008) (abcam; ab32101; 1:5000), anti-Bmi1 (Arigo; ARG55885; 1:1000), anti-survivin (Proteintech; 10508-1-AP; 1:1000), anti-GAPDH (abcam; ab8245; 1:10,000).

Techniques: Transfection, Cell Culture, Staining, Flow Cytometry, Isolation, Western Blot, Software