Journal: FEBS Open Bio

Article Title: Liraglutide enhances the effect of checkpoint blockade in lung and liver cancers through the inhibition of neutrophil extracellular traps

doi: 10.1002/2211-5463.13499

Figure Lengend Snippet: Liraglutide inhibits tumor progression through neutrophils. Measure blood cells in (A) LLC and (B) Hepa1‐6 cells bearing C57BL/6 mice using complete blood count. Concentrations of circulating (C) GM‐CSF, (D) VEGF‐A, (E) MMP‐9, (F) myeloperoxidase, and (G) elastase in mice were measured using ELISA. (H) Circulating dsDNA was purified and measured using an ultramicrospectrophotometer. Statistical significance was determined with t ‐test. N = 6 mice for every group. Nonparametric tests were utilized to assess statistical significance between different treatment groups. * P < 0.05, ** P < 0.01, *** P < 0.001. Error bars represent SD.

Article Snippet: Murine Lewis lung cancer cell LLC and liver cancer cell Hepa1‐6 were cultured in Dulbecco's Modified Eagle's Medium containing 10% FBS and 1% streptomycin–penicillin (10 000 U·mL −1 ) (all: Corning, New York, NY, USA) in a humidified incubator at 37 °C containing 5% CO 2 .

Techniques: Enzyme-linked Immunosorbent Assay, Purification

Journal: FEBS Open Bio

Article Title: Liraglutide enhances the effect of checkpoint blockade in lung and liver cancers through the inhibition of neutrophil extracellular traps

doi: 10.1002/2211-5463.13499

Figure Lengend Snippet: Liraglutide reduced tumor‐infiltrated NETs. The LLC and Hepa1‐6 cells bearing C57BL/6 mice were treated using PBS or liraglutide, and neutrophils were enriched for in vitro study. Representative images of tumor‐derived NETs from (A) LLC and (C) Hepa1‐6 cells bearing C57BL/6 mice. Blue is nuclear stained by Hoechst 33342. Green is DNA stained by SYTOX Green. Quantification of tumor‐derived NETs from (B) LLC and (D) MC38 cells bearing C57BL/6 mice were conducted using a fluorescence microplate reader ( n = 3). The tumor infiltrated neutrophils from (E) LLC and (G) Hepa1‐6 cells bearing C57BL/6 mice were stained with total reactive oxygen species (ROS) assay kit, ROS production was analyzed by flow cytometry, and the green fluorescence of ROS was detected using FITC channel. Mean fluorescence intensity (MFI) of ROS in tumor‐derived neutrophils from (F) LLC and (H) MC38 cells bearing C57BL/6 mice were conducted using a fluorescence microplate reader ( n = 3). Nonparametric tests were utilized to assess statistical significance between different treatment groups. * P < 0.05, *** P < 0.001. Error bars represent SD. Scale bar: 400 μm (A, C).

Article Snippet: Murine Lewis lung cancer cell LLC and liver cancer cell Hepa1‐6 were cultured in Dulbecco's Modified Eagle's Medium containing 10% FBS and 1% streptomycin–penicillin (10 000 U·mL −1 ) (all: Corning, New York, NY, USA) in a humidified incubator at 37 °C containing 5% CO 2 .

Techniques: In Vitro, Derivative Assay, Staining, Fluorescence, ROS Assay, Flow Cytometry

Journal: FEBS Open Bio

Article Title: Liraglutide enhances the effect of checkpoint blockade in lung and liver cancers through the inhibition of neutrophil extracellular traps

doi: 10.1002/2211-5463.13499

Figure Lengend Snippet: Liraglutide enhanced the antitumor efficacy of PD‐1 blockade. Tumor volumes were monitored in (A) LLC or (B) Hepa1‐6 allograft mice treated by IgG isotype control (isotype), anti‐PD‐1 mAb (αPD‐1), liraglutide, or liraglutide and anti‐PD‐1 mAb (liraglutide+αPD‐1). (C) LLC and (D) Hepa1‐6 tumor volumes for the NETs‐removed C57BL/6 mice. NETs were removed using DNase I (5 mg·kg −1 ). (E) LLC and (F) Hepa1‐6 tumor volumes for the NETs‐removed C57BL/6 mice accepting isotype, αPD‐1, liraglutide, or liraglutide+αPD‐1 treatments. NETs were removed using DNase I (5 mg·kg −1 ). Tumor volume = 0.5 × length × width 2 . Statistical significance was determined with two‐way ANOVA tests. N = 12 mice for every group. Nonparametric tests were utilized to assess statistical significance between different treatment groups. ** P < 0.01, *** P < 0.001. Error bars represent SD.

Article Snippet: Murine Lewis lung cancer cell LLC and liver cancer cell Hepa1‐6 were cultured in Dulbecco's Modified Eagle's Medium containing 10% FBS and 1% streptomycin–penicillin (10 000 U·mL −1 ) (all: Corning, New York, NY, USA) in a humidified incubator at 37 °C containing 5% CO 2 .

Techniques: Control

Journal: FEBS Open Bio

Article Title: Liraglutide enhances the effect of checkpoint blockade in lung and liver cancers through the inhibition of neutrophil extracellular traps

doi: 10.1002/2211-5463.13499

Figure Lengend Snippet: Long‐term immune responses induced by the combination therapy rejected rechallenge. The measurement of IFN‐γ concentration in the culture media of CD8 + T cells from (A, B) lymph node, (C, D) spleen, and (E, F) tumor tissues in LLC or Hepa1‐6 allograft mice after 7 days of treatment. N = 12 mice for every group. (G) LLC or (H) Hepa1‐6 allograft C57BL/6 mice were treated with STR. Mice pre‐treated with PBS, or the combination therapy received tumor rechallenge by administration of respective cell lines. N = 6 mice for every group. Tumor volume = 0.5 × length × width 2 . Statistical significance was determined with two‐tailed t ‐test. Nonparametric tests were utilized to assess statistical significance between different treatment groups. * P < 0.05, ** P < 0.01, *** P < 0.001. Error bars represent SD.

Article Snippet: Murine Lewis lung cancer cell LLC and liver cancer cell Hepa1‐6 were cultured in Dulbecco's Modified Eagle's Medium containing 10% FBS and 1% streptomycin–penicillin (10 000 U·mL −1 ) (all: Corning, New York, NY, USA) in a humidified incubator at 37 °C containing 5% CO 2 .

Techniques: Concentration Assay, Two Tailed Test

Journal: BMC Complementary and Alternative Medicine

Article Title: Xiao-Qing-Long-Tang shows preventive effect of asthma in an allergic asthma mouse model through neurotrophin regulation

doi: 10.1186/1472-6882-13-220

Figure Lengend Snippet: XQLT inhibited NGF, BDNF, TARC and p75NTR expression in LA4 cells stimulated with Der p. Pre-treatment (XP) and co-treatment (XD) with XQLT in Der p-stimulated cells and merely XQLT treated cells (XC) were used to assess the effects of XQLT on neurotrophin and associated receptor. (A) The supernatant was collected and analyzed by ELISA to obtain NGF, BDNF and TARC levels. Student’s t test was performed versus Group D. Schematic of cell experiment groups is also shown under those ELISA data. (B) Whole cell extracts were used for western blotting with anti-p75NTR antibody or anti-NGF antibody (commercial antibody also recognizes pro-NGF). Protein expression level was assessed as western band density detected by densitometry and was depicted as proportional bar plots (negative group level set as 1). Student’s t test was performed versus Group D. All the experiments in (A) and (B) were repeated for at least three times. The values shown are means ± SD. * means p < 0.05, ** means p < 0.01, *** means p < 0.005 in (A) and (B) .

Article Snippet: LA4 (murine lung adenoma) cell line was purchased from the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan; BCRC60239 derived from ATCC: CCL-196).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Western Blot