Journal: International Journal of Molecular Sciences

Article Title: Mesenchymal Stem Cells and Formyl Peptide Receptor 2 Activity in Hyperoxia-Induced Lung Injury in Newborn Mice

doi: 10.3390/ijms231810604

Figure Lengend Snippet: The effects of MSCs and FPR2 inhibitor, WRW4, on RAW264.7 cells’ inflammatory reaction. ( A ) Levels of FPR2 normalized to GAPDH loading control (Full length Western blots are shown in ) and ( B ) inflammatory cytokines, such as IL-1α and TNF-α, were measured in RAW264.7 cells, 12 h after H 2 O 2 (100 μM) induction, with or without WRW4 (10 μM) and MSC treatments (RAW264.7 cells: MSC ratio of 5:1). * p < 0.05 vs. control group. # p < 0.05 vs. H 2 O 2 - treated group. ( C ) Viabilities of L2 lung epithelial cells and HULEC-5a lung endothelial cells were measured after incubation in conditioned media (CM) of H 2 O 2 -induced RAW264.7 cells (100 μM of H 2 O 2 for 12 h), treated with WRW4 (10 μM) and MSCs (RAW264.7 cells: MSC ratio of 5:1), or non-treated. Data are given as mean ± SEM. * p < 0.05 vs. normal control group. † p < 0.05 vs. normoxia with WRW4-treated group. # p < 0.05 vs. H 2 O 2 control group.

Article Snippet: The effects of MSCs and WRW4, a FPR2 antagonist, on the viability of lung epithelial cell lines (L2 cells, Korean Cell Line Bank, Seoul National University College of Medicine, Seoul, Korea) and lung endothelial cell lines (HULEC-5a cells, American Type Culture Collection, Manassas, VA, USA) were measured using a Cell Counting Kit-8 assay (CCK-8; Dojindo, Kumamoto, Japan).

Techniques: Control, Western Blot, Incubation

Journal: Scientific Reports

Article Title: The c-Jun and JunB transcription factors facilitate the transit of classical Hodgkin lymphoma tumour cells through G 1

doi: 10.1038/s41598-018-34199-9

Figure Lengend Snippet: shRNA–mediated knock-down of JunB in ALK+ ALCL cell lines results in reduced proliferation. Western blots showing JunB levels ( A – C ) and growth curves ( D – F ) of the indicated cell lines expressing either control or JunB shRNA. Growth curves represent five experiments from three infections ( D ), five experiments from five infections ( E ), and three experiments from one infection ( F ). Note: JunB#6 and JunB#1 shRNAs overlap in their seed sequence and are not distinct. BrdU/7-AAD double staining of Karpas 299 ( G ) or SUP-M2 ( H ) cells expressing control or JunB shRNA. Results represent the average and standard deviation of three independent experiments from three infections for Karpas 299 cells, and eight independent experiments from six infections for SUP-M2 cells. ( I ) Bar graph showing the percentage of cells in S phase (% BrdU positive cells) observed when control shRNA or JunB shRNA–expressing Karpas 299 cells were transfected with plasmids expressing either EGFP-P2A or EGFP-P2A-FLAG-JunB. The results represent the average and standard deviation of four independent experiments. ( J ) Approximate doubling times and time spent in each stage of the cell cycle was determined for Karpas 299 and SUP-M2 cells expressing control or JunB shRNA. ( K ) Representative flow cytometry data and summary of Ki-67 expression within the G 0 /G 1 population of SUP-M2 cells expressing control or JunB shRNA. The summary represents the average and standard deviation of three independent experiments from two separate infections. P values were obtained by performing independent, two-tailed t tests comparing the JunB knock-down to control shRNA–expressing cells ( D – F and K ) or between JunB knock-down cells with or without JunB cDNA ( I ). ANOVA with Tukey’s post hoc test was performed in ( I ). * P < 0.05, ** P < 0.01, *** P < 0.001. Molecular mass markers (in kDa) are indicated to the left of western blots.

Article Snippet: The UCONN-L2 ALK+ ALCL cell line was obtained from Dr. Raymond Lai (University of Alberta; Edmonton, AB, Canada).

Techniques: shRNA, Knockdown, Western Blot, Expressing, Control, Infection, Sequencing, Double Staining, Standard Deviation, Transfection, Flow Cytometry, Two Tailed Test

Journal: Scientific Reports

Article Title: The c-Jun and JunB transcription factors facilitate the transit of classical Hodgkin lymphoma tumour cells through G 1

doi: 10.1038/s41598-018-34199-9

Figure Lengend Snippet: shRNA–mediated knock-down of c-Jun in ALK+ ALCL cell lines does not impair proliferation. Western blots showing c-Jun levels ( A , C , and E ) and growth curves ( B , D , and F ) of the indicated cell lines expressing either control or c-Jun shRNA. Growth curves represent four experiments from four infections (c-Jun#1) and five experiments from three experiments (c-Jun#5) ( B ), three experiments from three infections ( D ), and three experiments from one infection ( F ). Although, not included in the growth curve shown, c-Jun knock-down in UCONN-L2 had no effect on proliferation in other infections. ( G ) The percentage of cells in each stage of the cell cycle was measured by BrdU/7-AAD double staining for SUP-M2 cells expressing either control or c-Jun shRNA. The results represent three experiments from three infections. No statistically significant differences in growth rate or cell cycle distribution were observed between any c-Jun knock-down cells and respective control shRNA–expressing cells. Molecular mass markers (in kDa) are indicated to the left of western blots.

Article Snippet: The UCONN-L2 ALK+ ALCL cell line was obtained from Dr. Raymond Lai (University of Alberta; Edmonton, AB, Canada).

Techniques: shRNA, Knockdown, Western Blot, Expressing, Control, Infection, Double Staining

Journal: Biomedicines

Article Title: Mesenchymal Stem Cells Reduce the Extracellular Mitochondrial DNA-Mediated TLR9 Activation in Neonatal Hyperoxia-Induced Lung Injury

doi: 10.3390/biomedicines12030686

Figure Lengend Snippet: Cell viability and the level of extracellular mtDNA in H 2 O 2 -induced L2 rat lung epithelial cells. ( a ) Cell viability decreased in an H 2 O 2 concentration-dependent manner in the culture media. ( b ) Densitometric analysis of extracellular mtDNA and the representative PCR gel images. The full-length PCR gel is shown in . Data are expressed as mean ± SEM ( n = 10 per group). * p < 0.05 vs. normoxic control, # p < 0.05 vs. H 2 O 2 50 μM, $ p < 0.05 vs. 100 μM. ( c ) Cell viability of nontreated normoxic control, 100 μM H 2 O 2 induction, and 100 μM H 2 O 2 induction with MSC treatment (10:1 ratio). ( d ) Densitometric analysis of extracellular mtDNA and the representative PCR gel image of nontreated normoxic control, 100 μM H 2 O 2 induction, and 100 μM H 2 O 2 induction with MSC treatment (10:1 ratio). The full-length PCR gel is shown in . Data are expressed as mean ± SEM ( n = 10 per group). * p < 0.05 vs. normoxic control, # p < 0.05 vs. H 2 O 2 .

Article Snippet: The rat lung epithelial L2 cell line was purchased from the Korean Cell Line Bank (Seoul, Republic of Korea).

Techniques: Concentration Assay, Control