Journal: Neural Regeneration Research

Article Title: The circ_0002538/miR-138-5p/plasmolipin axis regulates Schwann cell migration and myelination in diabetic peripheral neuropathy

doi: 10.4103/1673-5374.355979

Figure Lengend Snippet: Characterization of circ_0002538 and its function in SCs. (A) Hierarchical clustering analyses of DEcircRNAs ( n = 3). (B, C) RT-PCR verified five circRNAs with upregulated expression and six circRNAs with downregulated expression, and the results were consistent with the RNA-seq data ( n = 12). The red dotted box highlights the circRNA of interest. Y -axis: Fold changes in circRNA expression compared with the non-diabetic group. * P < 0.05, ** P < 0.01, vs . non-diabetic group (independent-sample t-test). (D) Schematic diagram showing that circ_0002538 was formed by the circularization of KLHL8 exon 2. The red arrow represents the “head-to-tail” splicing site of circ_0002538, confirmed by Sanger sequencing. (E) We used divergent primers and convergent primers to amplify circ_0002538 in cDNA and gDNA. We used β-actin as a negative control. (F) circ_0002538 and KLHL8 mRNA in SCs were detected via RT-PCR after incubation with or without RNase R. Y -axis: fold changes in RNA expression compared with the mock group. *** P < 0.001, vs . mock group (independent-sample t -test). (G) circ_0002538 and KLHL8 mRNA levels were evaluated in the sh-circ_0002538-transfected SCs via RT-PCR. Y -axis: fold changes in RNA expression compared with the sh-NC group. *** P < 0.001, vs . sh-NC group. (H, I) The migrating number of SCs in the sh-circ_0002538 group was lower than that in the sh-NC group in the Transwell assays. ** P < 0.01, vs . sh-NC group (independent-sample t -test). Scale bars: 100 μm. All bar graphs represent the average of at least three independent replicates, and the error bars are the SD. cDNA: Complementary DNA; DPN: diabetic peripheral neuropathy; gDNA: genomic DNA; KLHL8: Kelch-like family member 8; sh-circ_0002538: short hairpin RNA for circ_0002538; sh-NC: normal control for short hairpin RNA.

Article Snippet: To detect the combination of circRNAs and miRNAs, we performed RNA pulldown assays with biotinylated probes according to the manufacturer’s protocol (MCE, Shanghai, China, Cat# HY-K0208).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, RNA Sequencing, Sequencing, Negative Control, Incubation, RNA Expression, Transfection, shRNA, Control

Journal: Scientific Reports

Article Title: Motility and chemotaxis of bacteria-driven microswimmers fabricated using antigen 43-mediated biotin display

doi: 10.1038/s41598-018-28102-9

Figure Lengend Snippet: Microparticle attachment and motility of the fabricated bacteriabots. Motile wild-type E. coli cells (WT) and non-motile Δ fliC cells carrying the recombinant Ag43-BAP and an inducible GFP construct were incubated for 20 min with streptavidin-coated 2.2-µm PMMA particles, at a mixing ratio of 1:30. ( A ) Images of particle-attached cells acquired via confocal laser scanning microscopy (top) or scanning electron microscopy (bottom), with scale bars being 2 µm and 1 µm, respectively. For better visualization, cells were elongated by inhibiting cell division using cephalexin to the growth medium for one hour before harvesting. ( B ) Corresponding quantification of the polar and non-polar cell-particle attachment of WT (for 786 attached cells) and Δ fliC (for 1115 attached cells) cells were analyzed via fluorescence microscopy. Statistical analysis was performed using a two-sample t -test with unequal sample size and unequal variance, with asterisk indicating P < 0.005. Error bars show SEM of three independent experiments. ( C ) Kinetics of particle attachment quantified using flow cytometry. Where indicated, CCCP was added during incubation to reduce cell motility. As a negative control, biotin was added in excess to inhibit the cell-particle attachment. Statistical analysis, performed using a two-sample t -test with unequal sample size and unequal variance, showed that difference between all datasets was highly significant ( P < 0.00001). Data are from six independent experiments. ( D ) Exemplary trajectories of bacteriabots with one, two and three attached WT E. coli cells, analyzed using time-lapse fluorescence microscopy. Acquisition time of depicted trajectories was 6.5 s (panel I), 7.4 s (panel II), 13 s (panel III), and 5.4 s (panel IV). Scale bar: 8 µm.

Article Snippet: The cell suspension was mixed in a ratio of 1:30 (in numbers) with 1.4-μm or 2.2-μm Red4 streptavidin PMMA particles (1.5 × 10 3 particles/μl; PolyAn, Berlin) and incubated for 20 min at room temperature.

Techniques: Recombinant, Construct, Incubation, Confocal Laser Scanning Microscopy, Electron Microscopy, Fluorescence, Microscopy, Flow Cytometry, Negative Control

Journal: Scientific Reports

Article Title: Motility and chemotaxis of bacteria-driven microswimmers fabricated using antigen 43-mediated biotin display

doi: 10.1038/s41598-018-28102-9

Figure Lengend Snippet: Motility of bacteriabots based on normal and elongated cells. ( A – E ) Distribution of swimming speed within populations of normal or elongated E. coli cells, with or without PMMA microparticles, as indicated. Average swimming speed of individual cells was determined based on 2D trajectories recorded using fluorescence microscopy, as described in Methods. Data are from seven or more independent experiments. Numbers of analyzed trajectories were 109192, 30724, 44698, 10601, and 10744 for cells, cells with 2.2-µm particles, cells with 1.4-µm particles, elongated cells, and elongated cells with 1.4-µm particles, respectively ( F ) Examples of 2D trajectories of bacteriabots based on elongated E. coli cells. Scale bar: 8 µm. ( G ) Autocorrelation of direction of motion, calculated from the trajectory data as described in Methods.

Article Snippet: The cell suspension was mixed in a ratio of 1:30 (in numbers) with 1.4-μm or 2.2-μm Red4 streptavidin PMMA particles (1.5 × 10 3 particles/μl; PolyAn, Berlin) and incubated for 20 min at room temperature.

Techniques: Fluorescence, Microscopy