Journal: Veterinary Research

Article Title: The local immune response of mice after Helicobacter suis infection: strain differences and distinction with Helicobacter pylori

doi: 10.1186/1297-9716-43-75

Figure Lengend Snippet: Number of colonizing H. suis bacteria. Shown is the average number of H. suis bacteria/milligram tissue in the stomach of BALB/c and C57BL/6 mice 59 days after experimental infection.

Article Snippet: Seven-week-old, female specific-pathogen-free BALB/c and C57BL/6 mice, free of Helicobacter spp., were purchased from Harlan NL (Horst, The Netherlands).

Techniques: Bacteria, Infection

Journal: Veterinary Research

Article Title: The local immune response of mice after Helicobacter suis infection: strain differences and distinction with Helicobacter pylori

doi: 10.1186/1297-9716-43-75

Figure Lengend Snippet: General cytokine expression profile after experimental H. suis infection in BALB/c and C57BL/6 mice. Shown are the mean fold changes in mRNA expression of indicated cytokines in H. suis -infected BALB/c and C57BL/6 mice. The mean fold change in the relevant uninfected control groups is equal to 1. An * indicates a statistically significant difference compared to uninfected control mice ( p < 0.05).

Article Snippet: Seven-week-old, female specific-pathogen-free BALB/c and C57BL/6 mice, free of Helicobacter spp., were purchased from Harlan NL (Horst, The Netherlands).

Techniques: Expressing, Infection, Control

Journal: Veterinary Research

Article Title: The local immune response of mice after Helicobacter suis infection: strain differences and distinction with Helicobacter pylori

doi: 10.1186/1297-9716-43-75

Figure Lengend Snippet: Cytokine expression profiles for each group of animals infected with H. suis strains HS1-9 and H. pylori strains SS1 or pMSS1. Shown are the mean fold changes of mRNA expression in HS1-HS9- and SS1- or pMSS1-infected BALB/c and C57BL/6 mice for IL-1β ( A ), IL-4 ( B ), IL-6 ( C ), IL-10 ( D ) and IL-17 ( E ). The mean fold change in the relevant uninfected control groups is equal to 1. An * indicates a significant upregulation of mRNA expression compared to uninfected control mice ( p < 0.05). An ** indicates a significant upregulation of mRNA expression compared to uninfected control mice ( p < 0.0056, i.e. the new cut-off p value obtained after Bonferroni correction for multiple comparisons).

Article Snippet: Seven-week-old, female specific-pathogen-free BALB/c and C57BL/6 mice, free of Helicobacter spp., were purchased from Harlan NL (Horst, The Netherlands).

Techniques: Expressing, Infection, Control

Journal: Veterinary Research

Article Title: The local immune response of mice after Helicobacter suis infection: strain differences and distinction with Helicobacter pylori

doi: 10.1186/1297-9716-43-75

Figure Lengend Snippet: Correlation between cytokine expression and H. suis colonization. Shown are the correlation analyses between IL-6 ( A ), IL-17 ( B ), MIP-2 ( C ), IL-1β ( D ) and IL-10 ( E ) mRNA expression levels and the number of colonizing H. suis bacteria in stomachs of indicated mouse strains (BALB/c and C57BL/6). Correlation was measured by Spearman’s Rho (ρ).

Article Snippet: Seven-week-old, female specific-pathogen-free BALB/c and C57BL/6 mice, free of Helicobacter spp., were purchased from Harlan NL (Horst, The Netherlands).

Techniques: Expressing, Bacteria

Journal: Oncogene

Article Title: miR-4516 predicts poor prognosis and functions as a novel oncogene via targeting PTPN14 in human glioblastoma

doi: 10.1038/s41388-018-0601-9

Figure Lengend Snippet: (A-B) Cell proliferation assay for U87MGvIII-luc/GBM30-luc cells stably transfected with lentivirus control or lentivirus miR-4516. The cell proliferation rates at 72 hours were measured by methylene blue. n=5, **P<0.01 (C) U87MG/EGFRvIII-luc/NC cells and U87MG/EGFRvIII-luc/miR-4516 cells (100 cells/well) were seeded into 6-well plates for colony formation assay and cultured for ~2 weeks to allow colony formation. The number of detectable colonies was counted. n=3, **P<0.0. (D) GBM30-luc/NC cells and GBM30-luc/miR-4516 cells (1000 cells/well) were seeded into 6-well plates for sphere formation assay and cultured for ~2 weeks. The number of detectable colonies was counted. n=3, **P<0.01. (E) Luminescent imaging for mice with intracranial tumor formed from GBM30-luc/NC and GBM30-luc/miR-4516 cells on day 10 after implanting (left). Quantification of luminescence signal intensity (right). Student t test was carried out for statistical analysis (mean ±SEM, n=9). (F) Kaplan-Meier survival analysis shows that overall survival of U87MGvIII-luc or GBM30-luc cells with stable overexpression of miR-4516 in comparison with miRNA-NC (n=9) (G) Representative images of strongly positive or weakly positive PTPN14 in GBM30-luc/NC and GBM30-luc/miR-4516 tissue are presented. Arrow head identifies a blood vessel with positive PTPN14 expression in endothelial cells as opposed to lack of expression on neoplastic cells. Magnification: ×400. (H) Tumors were formalin-fixed, paraffin-embedded, sliced and stained with H&E. Representative images of each group are presented. Magnification: left: ×200, right: ×400. Cell proliferation was determined by Ki-67 using immunohistochemistry. Magnification: ×400. (I) The PTPN14 protein in GBM30-luc/NC and GBM30-luc/miR-4516 tissues displayed an inverse correlation with the expression of miR-4516.

Article Snippet: For siRNA transfection, a specific PTPN14 siRNA (50 nmol/L) (OriGene) and a negative control were used. pcDNA3/ PTPN14 plasmid containing PTPN14 open reading frame (plasmid #61003) was purchased from Addgene (Cambridge, MA).

Techniques: Proliferation Assay, Stable Transfection, Transfection, Control, Colony Assay, Cell Culture, Tube Formation Assay, Imaging, Over Expression, Comparison, Expressing, Formalin-fixed Paraffin-Embedded, Staining, Immunohistochemistry

Journal: Oncogene

Article Title: miR-4516 predicts poor prognosis and functions as a novel oncogene via targeting PTPN14 in human glioblastoma

doi: 10.1038/s41388-018-0601-9

Figure Lengend Snippet: (A-C) U87MG, LN18, and GBM30 cells were transfected with either control (pcDNA3) or pcDNA3/PTPN14 plasmid. Forty-eight hours post-transfection, western blotting was performed to determine the protein expression of PTPN14, pYAP, YAP, and GAPDH using respective antibodies. The intensity of each band was quantified using ImageJ and normalized to GAPDH and then to pcDNA3-transfected cells. (D-F) U87MG, LN18, and GBM30 cells were transiently transfected with either miRNA negative control (NC) or miR-4516 mimic. After 48 hours, western blotting was performed to determine the protein level of PTPN14, pYAP, YAP, and GAPDH using respective antibodies. The intensity of each band was quantified using ImageJ and normalized to GAPDH and then to NC-transfected cells.

Article Snippet: For siRNA transfection, a specific PTPN14 siRNA (50 nmol/L) (OriGene) and a negative control were used. pcDNA3/ PTPN14 plasmid containing PTPN14 open reading frame (plasmid #61003) was purchased from Addgene (Cambridge, MA).

Techniques: Transfection, Control, Plasmid Preparation, Western Blot, Expressing, Negative Control

Journal: Oncogene

Article Title: miR-4516 predicts poor prognosis and functions as a novel oncogene via targeting PTPN14 in human glioblastoma

doi: 10.1038/s41388-018-0601-9

Figure Lengend Snippet: (A) Luciferase reporter containing 3 putative miR-4516 targeting sequences in the 3’UTR of PTPN14 (PTPN14 3’UTR wild-type) and mutated versions, containing 3 altered nucleotides (red). (B) LN229, U87MG, LN18, and GBM30 cells were co-transfected with the luciferase reporter constructs containing wild type (wt) or mutated PTPN14 3’UTR (mut) and NC or miR4516 mimic into. n=3, ***P<0.001. 72 hours post-transfection luciferase activity was measured and normalized. (C-D) qPCR and Western blotting to measure PTPN14 mRNA and protein levels in LN229, U87MG, LN18, and GBM30 cells transfected with NC or miR-4516 mimic. 72 hours post-transfection, cells were harvested for RNA and protein extractions. n=3, **P<0.01, ***P<0.001. (E) Immunohistochemical staining of PTPN14 on TMA showed the differential expression of PTPN14 in GBM tissues with low or high miR-4516 expression. Magnification: ×100 and ×200. (F) An inverse relationship between miR-4516 expression level and the PTPN14 protein level in 56 GBM tissues. P=0.012. Score 0/1 was considered as low expression of PTPN14, whereas score 2/3 together as high expression of PTPN14.

Article Snippet: For siRNA transfection, a specific PTPN14 siRNA (50 nmol/L) (OriGene) and a negative control were used. pcDNA3/ PTPN14 plasmid containing PTPN14 open reading frame (plasmid #61003) was purchased from Addgene (Cambridge, MA).

Techniques: Luciferase, Transfection, Construct, Activity Assay, Western Blot, Immunohistochemical staining, Staining, Quantitative Proteomics, Expressing

Journal: Oncogene

Article Title: miR-4516 predicts poor prognosis and functions as a novel oncogene via targeting PTPN14 in human glioblastoma

doi: 10.1038/s41388-018-0601-9

Figure Lengend Snippet: (A) Western blot analysis for PTPN14 in U87MG/LN229 cells transfected with siRNA control or PTPN14 siRNA for 48hrs. (B) Cell proliferation assay for U87MG/LN229 cells transiently transfected with PTPN14 siRNA. The cell proliferation rates were measured by methylene blue. n=5, **P<0.01. (C) Cell invasion assay for U87MG/LN229 cells transiently transfected with PTPN14 RNAi using matrigel transwell membranes. n=3, ***P<0.001. (D-F) U87MG/LN229 cells were transiently transfected with NC or miR-4516 inhibitor alone, or simultaneously with PTPN14 RNAi. The protein expression of PTPN14 was determined using westen blotting (D). Cell proliferation at 72 hours was determined using methylene blue (E). n=5, **, P < 0.01. The matrigel transwell invasion assay was carried out to quantify the invaded cells (F). n=3, ***, P < 0.001.

Article Snippet: For siRNA transfection, a specific PTPN14 siRNA (50 nmol/L) (OriGene) and a negative control were used. pcDNA3/ PTPN14 plasmid containing PTPN14 open reading frame (plasmid #61003) was purchased from Addgene (Cambridge, MA).

Techniques: Western Blot, Transfection, Control, Proliferation Assay, Invasion Assay, Expressing, Transwell Invasion Assay

Journal: Redox Biology

Article Title: A novel role of KEAP1/PGAM5 complex: ROS sensor for inducing mitophagy

doi: 10.1016/j.redox.2021.102186

Figure Lengend Snippet: PGAM5 interferes with PINK1 processing independently of its phosphatase activity. A. PGAM5 mutant and isoform lacking phosphatase activity induce Parkin translocation. PC6 cells were transfected with Parkin-EYFP and PGAM5 wt, PGAM5 F244D or short PGAM5 isoform (isoform 2, UniProt #Q96HS1-2). ****P < 0.0001, n = 6 dishes, 20 fields per dish, Welch's ANOVA followed by Dunnett's T3 multiple comparisons test. B. PGAM5 mutant lacking PARL/OMA1 cleaving site does not induce Parkin translocation. PC6 cells were transfected with Parkin-EYFP and PGAM5 or PGAM5 S24F. ****P < 0.0001, n = 6 dishes, 20 fields per dish, Welch's ANOVA followed by Dunnett's T3 multiple comparisons test. C. TOMM7 overexpression increases the fraction of full-length PGAM5 in HEK293 cells expressing PGAM5-flag and TOMM7-myc. D. TOMM7 overexpression protects partially against PGAM5 induced Parkin translocation. PC6 cells were transfected with Parkin-EYFP, PGAM5 wt or/and TOMM7. ****P < 0.0001, n = 9 dishes, 20 fields per dish, Welch's ANOVA followed by Dunnett's T3 multiple comparisons test. E. OMA1 and PARL protect partially against PGAM5 induced Parkin translocation. PC6 cells were transfected with Parkin-EYFP, PGAM5 wt and PARL or OMA1. ****P < 0.0001, n = 6 dishes, 20 fields per dish, One-way ANOVA followed by Sidak's multiple comparisons test. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: PARL-FLAG , Addgene , Cat# 13639.

Techniques: Activity Assay, Mutagenesis, Translocation Assay, Transfection, Over Expression, Expressing

Journal: Redox Biology

Article Title: A novel role of KEAP1/PGAM5 complex: ROS sensor for inducing mitophagy

doi: 10.1016/j.redox.2021.102186

Figure Lengend Snippet:

Article Snippet: PARL-FLAG , Addgene , Cat# 13639.

Techniques: Recombinant, Lysis, Extraction, Blocking Assay, Isolation, DC Protein Assay, Caspase-Glo Assay, shRNA, Generated, Plasmid Preparation, Synthesized, Variant Assay

Journal: iScience

Article Title: Friendly fungi: Tropical insect families form partnerships with intracellular fungi related to pathogens

doi: 10.1016/j.isci.2024.110674

Figure Lengend Snippet:

Article Snippet: Sordaria fimicola , Carolina Biological Supply , Cat# 156290.

Techniques: Sequencing, Recombinant, Saline, Plasmid Preparation, Wright Stain, Picogreen Assay, Negative Control, Software