Journal: Scientific Reports

Article Title: DF2726A, a new IL-8 signalling inhibitor, is able to counteract chemotherapy-induced neuropathic pain

doi: 10.1038/s41598-019-48231-z

Figure Lengend Snippet: Western blotting analyisis for microglia marker Iba-1 and for astrocyte GFAP in control, OXA and (Oxa) + DF2726A rats. In the bottom, TRPA1 and TRPM8 receptors in in control, OXA and (Oxa) + DF2726A rats, evalutated by western blotting. Administration of DF2726A (Oxa + DF, light grey triangles) was able to significantly reduce reactive microglia and astrocytes as well as to reduce pain receptors. ***P < 0.001, **P < 0.01 and *P < 0.05 vs respective CTR group; +++ P < 0.001; ++ P < 0.01 and + P < 0.05 vs the respective Oxa + Saline group. 2-way repeated-measures ANOVA and Bonferroni test. Data are expressed as mean ± SEM; n 10 per group. ANOVA, analysis of variance.

Article Snippet: The filter was then blocked with 1 × PBS and 3% non-fat dried milk for 40 min at room temperature and probed with anti-Glial Fibrillary Acidic Protein (GFAP) antibody (diluition 1.1000; cat.no.Z0334, Dako), anti-ionized calcium-binding adapter molecule 1 (Iba-1) antibody (dilution 1:1000; cat. no.019-19741, Wako), anti-transient receptor potential ankyrin 1 (TRPA1) antibody (diluition 1:1000; cat. no. NB110-40763, Novus Biologicals) and anti- Transient receptor potential M8 (TRPM8) antibody (diluition 1:10100; cat. no. NBP1-97311, Novus Biologicals) in 1 × PBS, 3% non-fat dried milk, and 0.1% Tween 20 at 4 °C overnight.

Techniques: Western Blot, Marker, Control, Saline

Journal: PLOS ONE

Article Title: hnRNP Q/SYNCRIP interacts with LIN28B and modulates the LIN28B/ let-7 axis in human hepatoma cells

doi: 10.1371/journal.pone.0304947

Figure Lengend Snippet: ( A ) hnRNP Q depletion significantly reduced the relative luciferase activities of Trim71 3’UTR 468-bearing luciferase while having no effect on luciferase-containing 3’UTRs without a LCS. The psiCHECK2 vector carrying either the 468 bp TRIM71 3’UTR or the 424-bp 3’UTR with LCSs-deleted were transfected into control knockdown (black bars) or hnRNP Q knockdown (grey bars) HEK293 cells. For both the hnRNP Q knocked-down or control studies, luciferase activities were normalized to the samples with the psiCHECK2 empty vector. (B) hnRNP Q depletion significantly reduced relative luciferase activities of LCS-bearing luciferase in HEK293 cells. The psiCHECK2 vectors carrying 8X LCSs or 8X mutated LCSs were transfected into hnRNP Q knockdown (in grey) or control knockdown (in black) HEK293 cells for the assay. ( C ) hnRNP Q depletion significantly reduced the relative activities for luciferase constructs carrying either the 468 bp or 6,015 bp Trim71 3’UTRs in Huh7 cells. The pmirGLO vector containing either the 468 bp or 6,015 bp TRIM71 3’UTR were transfected into control knockdown (black bars) or hnRNP Q knockdown (grey bars) Huh7 cells for the assay. Results were plotted as foldchange (control = 1 for control knockdown or hnRNP Q knockdown samples for each set of luciferase activity tests) ± S.D. (D) Cell proliferation of Huh7 cells infected with lentiviral constructs containing sh_LacZ, sh_ hnRNP Q#1, or sh_ hnRNP Q#2 was determined by the MTT assay at the indicated time points. For = 4; = 3. n.s., not significant. * P < 0.05, ** P < 0.01, *** P < 0.001 using an unpaired two-tailed Student’s t -test. (E) Mantel-Cox survival curves comparing the effects of hnRNP Q expression in liver cancer.

Article Snippet: The TRIM71 3’ UTRs were also cloned to another dual luciferase plasmid, pmirGLO by using the Xho I and Not I restriction sites. psiCHECK2 plasmids containing eight tandem repeats of let-7 target or mutant sequences were obtained from Addgene (#20931) [ ].

Techniques: Luciferase, Plasmid Preparation, Transfection, Control, Knockdown, Construct, Activity Assay, Infection, MTT Assay, Two Tailed Test, Expressing

Journal: Biotechnology and bioengineering

Article Title: Microparticles enhance the formation of seven major classes of natural products in native and metabolically engineered actinobacteria through accelerated morphological development.

doi: 10.1002/bit.27818

Figure Lengend Snippet: Figure 7: Impact of talc microparticles on nine major classes of natural products across different actinobacterial families. RiPP-type cinnamycins using S. albus pCinCatInt (Lopatniuk et al., 2017), RiPP-type bottromycins using S. lividans TK24 DG2-Km-P41hyg+ (Horbal et al., 2018), alkaloid-type undecylprodigiosin using S. lividans TK24 DG2-Km-P41hyg+ (Horbal et al., 2018), angucyclinone-type simocyclinones using K. sp. (Bilyk et al., 2016), polyketide-type pamamycins using S. albus J1074/R2 (Rebets et al., 2015), polyketide-type alpiniamides using S. sp. IB2014/011-12 (Paulus et al., 2018), glycopeptide-type vancomycin using A. japonicum DSM 44213 (Stegmann et al., 2014), glycopeptide-type teicoplanin using A. teichomyceticus ATCC 31121 (Horbal et al., 2012), nybomycin using S. albus subsp. chlorinus NRRL B-24108 (Rodriguez Estevez et al., 2018), tetracycline-type oxytetracycline using S. rimosus ATCC 10970 (Pethick et al., 2013), tetrahydroisochinoline-type perquinolines using S. sp. IB2014/016-6 (Rebets et al., 2019), and anthramycin-type usabamycins using S. albus subsp. chlorinus NRRL B- 24108 (nybomycin) (Rodriguez Estevez et al., 2018). All strains were incubated over five days using miniaturized microtiter plate cultures (1 mL) at different talc levels between 0 and 50 g L-1. The natural product levels were determined after solvent extraction using HPLC-ESI-MS and reflect final titers after five days. The colored circles given beside strain names indicate the type of producers studied: native (blue circle) and heterologous strains (orange circle). The colored squares denote the underlying control type for cluster expression: native (blue square) and synthetic regulation (orange square). n=3.

Article Snippet: Strain Natural product Medium Reference Streptomyces albus J1074/R2 Pamamycins SGG (Rebets et al., 2015) Streptomyces lividans TK24 DG2-Km-P41hyg+ Bottromycins Undecylprodigiosin SG (Horbal et al., 2018) Streptomyces rimosus ATCC 10970 Oxytetracycline GOTC ATCC Streptomyces albus pCinCatInt Cinnamycins TSB (Lopatniuk et al., 2017) Streptomyces albus subsp. chlorinus NRRL B24108 Nybomycin Usabamycins DNPM (Myronovskyi et al., 2020) Streptomyces sp. IB2014/011-12 Alpiniamides TSB (Paulus et al., 2018) Streptomyces sp. IB2014/016-6 Perquinolines TSB (Rebets et al., 2019) Amycolatopsis japonicum DSM 44213 Vancomycin TSB DSMZ Actinoplanes teichomyceticus ATCC 31121 Teicoplanin TSB ATCC Kitasatospora sp. DSM 102431 Simocyclinones TSB DSMZ This article is protected by copyright.

Techniques: Glycoproteomics, Incubation, Solvent, Extraction, Control, Expressing