Journal: Stem Cells (Dayton, Ohio)

Article Title: Antibacterial Effect of Human Mesenchymal Stem Cells Is Mediated in Part from Secretion of the Antimicrobial Peptide LL-37

doi: 10.1002/stem.544

Figure Lengend Snippet: Antimicrobial activity of MSC CM is mediated by LL-37 secretion. (A): Synthetic LL-37 showed significant antimicrobial activity against E. coli in a dose-dependent manner. Data are mean ± SD; *, p < .001 versus RPMI by analysis of variance (ANOVA; Bonferroni) n = 3. (B): Synthetic LL-37 displayed dose-dependent antimicrobial effect against P. aeruginosa. Data are mean ± SD; *, p < .001 versus RPMI by ANOVA (Bonferroni) n = 3. Preincubation of MSC CM with anti-LL-37 antibody (1 μ;g/ml), but not with mouse IgG (1 μg/ml), significantly reduced the antimicrobial effect of MSC CM against E. coli (C) and P. aeruginosa (D). Data are mean ± SD; *, p < .0002 versus MSC CM + anti-LL-37 by ANOVA (Bonferroni), n = 5–7. Abbreviations: CFU, colony-forming unit; MSC CM, mesenchymal stem cell conditioned medium; RPMI, RPMI-1640 medium.

Article Snippet: Mouse monoclonal antibody to human LL37/CAP18 clone 3D11 and mouse isotype IgG1 antibody were purchased from Hycult Biotechnology (Netherlands), goat Alexa-Fluor 488-labeled anti-mouse-IgG from Invitrogen, synthetic human LL-37 from AnaSpec (CA) and fetal bovine serum (FBS) from HyClone Laboratories Inc. (Utah).

Techniques: Activity Assay

Journal: Stem Cells (Dayton, Ohio)

Article Title: Antibacterial Effect of Human Mesenchymal Stem Cells Is Mediated in Part from Secretion of the Antimicrobial Peptide LL-37

doi: 10.1002/stem.544

Figure Lengend Snippet: Intratracheal administration of anti-LL 37 antibody reduced the therapeutic effect of MSC in E. coli pneumonia. Coadministration of MSC together with an anti-LL 37 neutralizing antibody (10 μg), but not with mouse IgG isotype antibody, inhibited the therapeutic effect of MSC in bacterial clearance in lung homogenates (A) and BAL fluid (B). Values are mean CFU ± SD; *, p < .002 versus MSC + anti-Ll-37 antibody treated mice; √, p < .005 versus MSC + anti-LL-37 antibody-treated mice by analysis of variance (ANOVA; Bonferroni), n = 5. (C): MSC administration enhanced antimicrobial activity of mouse BAL. BAL samples were incubated with E. coli (105 CFU/ml) for 2 hours, and CFU growth was counted. Data are mean ± SD; *, p < .002 versus BAL of PBS-treated mice by ANOVA (Bonferroni), n = 8–9. Abbreviations: BAL, bronchoalveolar lavage; CFU, colony-forming unit; LH, lung homogenates; MSC, mesenchymal stem cell; PBS, phosphate buffered saline.

Article Snippet: Mouse monoclonal antibody to human LL37/CAP18 clone 3D11 and mouse isotype IgG1 antibody were purchased from Hycult Biotechnology (Netherlands), goat Alexa-Fluor 488-labeled anti-mouse-IgG from Invitrogen, synthetic human LL-37 from AnaSpec (CA) and fetal bovine serum (FBS) from HyClone Laboratories Inc. (Utah).

Techniques: Activity Assay, Incubation

Journal: PLOS One

Article Title: An in vitro tumor recurrence model based on platinum-resistant colon cancer cells as a research tool for studying cancer cell dormancy

doi: 10.1371/journal.pone.0333671

Figure Lengend Snippet: (A,B) LysoTracker-positive acidic lysosomal compartments in quiescent cells. Oxaliplatin-resistant HCT116 oxpl-R cells (A) and cisplatin-resistant HCT116 cspl-R cells (B) before (day 0) and after (day 8) platinum treatment (oxaliplatin or cisplatin, respectively), stained with LysoTracker Green™ and MitoTracker Orange™. Scale bars represent 50 µm. (C,D) Dynamics of autophagy marker expression in the in vitro cancer reсurrence model based on cisplatin-resistant colon cancer cells HCT116 cspl-R, analyzed by immunoblotting (C) and qPCR (D) . (C) Immunoblots probed with LC3, Beclin and Survivin antibodies. α-Tubulin was used as a loading control. Quantification values shown beneath each lane represent band intensity normalized to loading control and relative to day 0 control (set as 1.0). (D) Normalized expression of autophagy-related genes ( lc3, beclin ) after cisplatin exposure (days 0-33), relative to day 0. Expression of the GAPDH gene served as the endogenous control. Data represent biological triplicate experiments and are displayed as mean ± SEM.

Article Snippet: The primary antibodies used were against LC3 #4108 RRID: AB_2137703, Beclin #3738 RRID:AB_490837, Survivin #2808 RRID:AB_2063948 (Cell Signaling, USA), Cyclin A sc-751 RRID:AB_631329, and p27 (C-19) sc-528 RRID:AB_632129 (Santa Cruz Biotechnology, USA) with ɑ-Tubulin sc-32293 RRID:AB_628412 (Santa Cruz Biotechnology, USA) serving as a loading control.

Techniques: Staining, Marker, Expressing, In Vitro, Western Blot, Control

Journal: Matrix biology : journal of the International Society for Matrix Biology

Article Title: Elevated TGFβ signaling contributes to ocular anterior segment dysgenesis in Col4a1 mutant mice

doi: 10.1016/j.matbio.2022.05.001

Figure Lengend Snippet: (A) Schematic illustration of the 1D11 administration paradigm for histological analysis. (B) Representative images of H&E stained sections (left) showing whole eyes (top panels) and corneas (lower panels) from E18.5 mice and quantification graph (right) showing increased corneal stromal thickness in Col4a1+/G1344D mice treated with 1D11 compared to those that received the control IgG1 antibody. Scale bars = 200 μm (top) and 50 μm (bottom). n = 20 and 24 corneas from IgG1- and 1D11-treated Col4a1+/+ mice, and 24 and 30 corneas from IgG1- and 1D11-treated Col4a1+/G1344D mice, respectively. (C) Schematic illustration of the 1D11 administration paradigm for qPCR analyses. (D) qPCR analyses revealed that 1D11 treatment partially prevented the increased expression of TGFβ target genes in anterior segments from P0 Col4a1+/G1344D mice compared to their IgG1-treated counterparts. n = 5–6 samples per genotype. Data are presented as mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001; and ****p < 0.0001, two-way ANOVA and Tukey’s multiple comparison test.

Article Snippet: Timed-pregnant B6 females crossed with Col4a1 +/ G1344D males were injected intraperitoneally with the 1D11 pan-TGFβ neutralizing antibody (clone 1D11.16.8, BioXCell, West Lebanon, NH) or IgG1 isotype control antibody (clone MOPC-21, BioXCell) diluted in inVivoPure Dilution Buffer (pH 7.0, BioXCell) (20 mg/kg) every other day from E8.5 to E16.5 and animals were harvested at E18.5 or P0 for histological and molecular analyses, respectively.

Techniques: Staining, Control, Expressing, Comparison