Journal: Cell Reports Medicine

Article Title: Mechanical confinement promotes heat resistance of hepatocellular carcinoma via SP1/IL4I1/AHR axis

doi: 10.1016/j.xcrm.2023.101128

Figure Lengend Snippet: 3D bioprinted GCL construct (A) The schematic representation of the 3D thermal ablation model using the mixture of GCL hydrogel and HCC cell lines. (B) A photograph of a 3D bioprinted model with a grid-like structure. (C and D) Live/dead staining of MHCC-97H and quantifications in 3D model for days 1 and 7 (three biological replicates). (E) Schematic diagram and infrared imaging of orthotopic HCC tumor during thermal ablation for 30 s in BALB/C nude mice. (F) Picrosirius red staining of fibrillar collagen in the different regions of the tumor tissue after thermal ablation for 30 s. (G and H) The images and volume of GCL structure after different heat treatments for 15 min (three biological replicates). (I and J) Collagen hybridizing peptide staining of fibrillar collagen in the different regions of tumor tissue after thermal ablation for 30 s (five biological replicates). (K and L) SEM images of fibrillar collagen (red arrows) and quantifications in GCL structure after different heat treatments for 15 min (three biological replicates). (M) The stiffness of 3D (five biological replicates) and animal (10 biological replicates) models detected by nano-indentation under different temperatures. Mean ± standard deviation. Statistical analysis, one-way ANOVA for (D), (H), (J), (L), and (M, left), two-tailed Student’s t test for (M, right). n.s., not significant, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: A Picrosirius Red Staining Kit (ab150681, Abcam) was employed to reveal the fibrillar collagen of paraffinized tumor sections in the animal model of thermal ablation.

Techniques: Construct, Staining, Imaging, Standard Deviation, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: Mechanical confinement promotes heat resistance of hepatocellular carcinoma via SP1/IL4I1/AHR axis

doi: 10.1016/j.xcrm.2023.101128

Figure Lengend Snippet:

Article Snippet: A Picrosirius Red Staining Kit (ab150681, Abcam) was employed to reveal the fibrillar collagen of paraffinized tumor sections in the animal model of thermal ablation.

Techniques: Recombinant, Bicinchoninic Acid Protein Assay, Imaging, Staining, Transfection, Software, Microscopy, Fluorescence, Liquid Chromatography, Mass Spectrometry

Journal: Membranes

Article Title: Targeting the Structural Integrity of Extracellular Vesicles via Nano Electrospray Gas-Phase Electrophoretic Mobility Molecular Analysis (nES GEMMA)

doi: 10.3390/membranes12090872

Figure Lengend Snippet: Schematic drawing of the workflow for extracellular vesicle (EV) isolation and EV analysis performed in this research work. The following abbreviations are used: AA—ammonium acetate, nES GEMMA—nano electrospray gas-phase electrophoretic mobility molecular analyzer, nES DMA—nano electrospray differential mobility analyzer, NTA—nanoparticle tracking analysis, PBS—phosphate-buffered saline, RT—room temperature, SEC—size exclusion chromatography, US—ultrasonication.

Article Snippet: Subsequently, the single-charged, surface-dry particles were separated with a sheath flow of 8.0 L/min in a nano differential mobility analyzer (nDMA, model 3480C, TSI Inc.).

Techniques: Isolation, Gas Phase Electrophoretic Molecular Mobility Analysis, Size-exclusion Chromatography

Journal: PLoS ONE

Article Title: CD105 + -mesenchymal stem cells migrate into osteoarthritis joint: An animal model

doi: 10.1371/journal.pone.0188072

Figure Lengend Snippet: A) Selective enrichment of CD105 + expressing cells. Fluorescence-activated cell sorting (FACS) analysis of the subpopulations of CD105 + pre-sorted (on the top) and post-sorted (on the bottom) from synovial membrane. 105 after was subpopulation of CD105 + post-sorted (on the left bottom) and 105- after was subpopulation of CD105 - post-sorted (on the right bottom). B) Characterization by flow cytometry analysis of the CD105 + -MSC population using MSCs markers, CD44, CD105, CD90 and haematopoietic markers CD45 and CD34. C) Flow cytometry of subpopulation of CD105 + -MSC labelled with oxacarbocyanine (DiO) and octadecyl (C18) indocarbocyanine (DiI) respectively. D) Characterization of GFP-CD105 + -MSCs used to validate previous data obtained using DiO- CD105 + -MSCs and DiI-CD105 + -MSCs in animal model. Fluorescence-activated cell sorting (FACS) analysis of the MSCs populations (on the left), GFP-MSCs (on the middle) and post-sorted for CD105 + (on the right) from synovial membrane. Representative image of GFP-CD105 + -MSCs in a plate before injection, bar represents 20 μm.

Article Snippet: The primary antibodies used were mouse anti-human CD34 (1:20 DakoCytomation, Barcelone, SP), FITC mouse anti-human CD45 (1:20), FITC mouse anti-human CD105 (1:100 from Serotec, Bavaria, GER), FITC mouse anti-human CD44 (1:100 from Serotec, Bavaria, GER) and PE-Cy5-conjugated mouse anti-human CD90 (1:20 from BD Pharmagen, Madrid, SP).

Techniques: Expressing, Fluorescence, FACS, Membrane, Flow Cytometry, Animal Model, Injection

Journal: PLoS ONE

Article Title: CD105 + -mesenchymal stem cells migrate into osteoarthritis joint: An animal model

doi: 10.1371/journal.pone.0188072

Figure Lengend Snippet: A) Co-localization of DiO-CD105 + -MSCs with antibodies against anti-CD105, anti-CD44 and anti-CD90 in crossed ligaments sections of left knee from animals injected with DiO-CD105 + -MSCs IV (magnification were 20x, 40x and 60x). Arrows point cells which co-localized for CD44 and CD90 antibodies. B) Histogram showed percentage of DiO positives signal normalized with DAPI signal from animals IA injected in front of animals IV injected. AnalySIS Image Processing was used. C) Quantitative analysis to determine levels of positive DiO fluorescence in front of DAPI signal was done by AnalySIS Image software from tissues where DiO-CD105 + -MSCs were found. D) Immunofluorescence analysis of sections from patella femoral grove and synovial membrane from the left knee at different magnifications (10x, 20x and 40x) from animals injected IA with DiO-CD105 + -MSCs. Arrows point the same group of cells which co-localized for CD105, CD44 and CD90 antibodies. E) Immunofluorescence analysis of sections from synovial membrane at 20x and 40x magnifications from right knee (up) and left knee (down). F) Histogram showed percentage of CD44, CD90 and CD105 positives signal normalized with DAPI signal from animals injected IA with DiO-CD105 + -MSCs. AnalySIS Image Processing was used. * P value less than 0.05 was considered statistically significant by Mann Whitney and Kruskall-Wallis.

Article Snippet: The primary antibodies used were mouse anti-human CD34 (1:20 DakoCytomation, Barcelone, SP), FITC mouse anti-human CD45 (1:20), FITC mouse anti-human CD105 (1:100 from Serotec, Bavaria, GER), FITC mouse anti-human CD44 (1:100 from Serotec, Bavaria, GER) and PE-Cy5-conjugated mouse anti-human CD90 (1:20 from BD Pharmagen, Madrid, SP).

Techniques: Injection, Fluorescence, Software, Immunofluorescence, Membrane, MANN-WHITNEY

Journal: Nutrition & Metabolism

Article Title: The role of lipid droplet formation in the protection of unsaturated fatty acids against palmitic acid induced lipotoxicity to rat insulin-producing cells

doi: 10.1186/s12986-016-0076-z

Figure Lengend Snippet: Gene expression analysis of perilipin 1 or 2 suppressed RINm5F and INS-1E cells. Perilipin 1 and 2 expression in RINm5F ( a , c ) and INS-1E ( b , d ) cells was stably suppressed by the shRNA technique after lentiviral transduction. Cells were incubated with 100 μM palmitic acid (PA), 100 μM oleic acid (OA), a combination of both, or 100 μM gondoic acid (GA) or without NEFAs (Con) for 24 h. Thereafter RNA was isolated and reverse transcribed into cDNA. Gene expression of perilipin 1 and perilipin 2 was measured by qRT-PCR and normalized to the reference genes PPIA , α-tubulin and β-actin using qBase + software (Biogazelle). Data are means ± SEM of n = 4. * p < 0.01 compared to control cells without NEFAs (ANOVA/Tukey Multiple Comparison Test). # p < 0.01 compared with control cells (ANOVA/Tukey Multiple Comparison Test)

Article Snippet: The following antibodies from Santa Cruz Biotechnology (Santa Cruz, CA, USA) were used: anti-Perilipin 1 (H-300, diluted 1:2000), anti-Perilipin 2 (H-80, diluted 1:200), and anti-β-actin (sc-1615, diluted 1:250).

Techniques: Expressing, Stable Transfection, shRNA, Transduction, Incubation, Isolation, Quantitative RT-PCR, Software

Journal: Nutrition & Metabolism

Article Title: The role of lipid droplet formation in the protection of unsaturated fatty acids against palmitic acid induced lipotoxicity to rat insulin-producing cells

doi: 10.1186/s12986-016-0076-z

Figure Lengend Snippet: Western blot analyses of perilipin 1 or 2 suppressed RINm5F and INS-1E cells. Perilipin 1 and 2 expression in RINm5F ( a , c ) and INS-1E ( b , d ) cells was stably suppressed by the shRNA technique after lentiviral transduction. Cells were incubated with 100 μM palmitic acid (PA), 100 μM oleic acid (OA), a combination of both, or 100 μM gondoic acid (GA) or without NEFAs (Con) for 24 h. Thereafter, 30 μg protein of total whole cell extracts were separated by a SDS-PAGE and electroblotted on nitrocellulose membranes. Specific protein detection was carried out with a primary anti-perilipin 1 or anti-perilipin 2 antibody and a horseradish peroxidase labelled anti-rabbit IGG secondary antibody. After visualization of the protein bands by chemiluminescence the perilipin 1 and 2 bands were quantified and normalized to β-actin bands. Shown are means of ± SEM of n = 3 independent immunoblots and one representative blot for each cell line. * p < 0.01 compared to control cells without NEFAs (ANOVA/Tukey Multiple Comparison Test). # p < 0.01 compared with control cells (ANOVA/Tukey Multiple Comparison Test)

Article Snippet: The following antibodies from Santa Cruz Biotechnology (Santa Cruz, CA, USA) were used: anti-Perilipin 1 (H-300, diluted 1:2000), anti-Perilipin 2 (H-80, diluted 1:200), and anti-β-actin (sc-1615, diluted 1:250).

Techniques: Western Blot, Expressing, Stable Transfection, shRNA, Transduction, Incubation, SDS Page