Journal: Cells

Article Title: New Phage-Derived Antibacterial Enzyme PolaR Targeting Rothia spp.

doi: 10.3390/cells12151997

Figure Lengend Snippet: Antibacterial activity of PolaR against Rothia spp. over time. ( a ) Rothia dentocariosa PCM 2348 (ATCC 14189), ( b ) Rothia dentocariosa PCM 2349 (ATCC 17931), ( c ) Rothia mucilaginosa PCM 2403 and ( d ) Rothia mucilaginosa PCM 2415 (ATCC 25296) were tested in the Sytox Green fluorometric assay with three different PolaR concentrations: 1, 25 and 100 µg/mL. Increasing fluorescence represents an increasing number of killed bacterial cells. All samples were tested in triplicate. The presented data represent one of three independently conducted experiments on three independently produced PolaR preparations.

Article Snippet: Figure S1: SDS-PAGE of the final preparation of PolaR., Figure S2: Antibacterial activity of PolaR against Rothia aeria PCM 2669 in time, Figure S3: Antibacterial activity of 150 μg/mL PolaR against Rothia mucilaginosa PCM 2415 (ATCC 25296) was tested over time in a turbidity reduction assay (TRA).

Techniques: Activity Assay, Fluorescence, Produced

Journal: Cells

Article Title: New Phage-Derived Antibacterial Enzyme PolaR Targeting Rothia spp.

doi: 10.3390/cells12151997

Figure Lengend Snippet: The effect of PolaR on Rothia mucilaginosa PCM 2415 (ATCC 25296) aggregates. ( a ) LIVE/DEAD staining of Rothia mucilaginosa PCM 2415 (ATCC 25296) after 2–4 h treatment with PBS or 150 µg/mL of PolaR. ( b ) Ratio of aggregate size in 2–4 h after PBS or 150 µg/mL PolaR treatment. PBS fold change is marked as a red dashed line (* p = 0.02).

Article Snippet: Figure S1: SDS-PAGE of the final preparation of PolaR., Figure S2: Antibacterial activity of PolaR against Rothia aeria PCM 2669 in time, Figure S3: Antibacterial activity of 150 μg/mL PolaR against Rothia mucilaginosa PCM 2415 (ATCC 25296) was tested over time in a turbidity reduction assay (TRA).

Techniques: Staining

Journal: Cells

Article Title: New Phage-Derived Antibacterial Enzyme PolaR Targeting Rothia spp.

doi: 10.3390/cells12151997

Figure Lengend Snippet: Antibacterial activity of 150 µg/mL PolaR against sonicated and non-sonicated Rothia mucilaginosa PCM 2415 (ATCC 25296) was tested over time in a fluorometric assay. Increasing fluorescence represents an increasing number of killed bacterial cells.

Article Snippet: Figure S1: SDS-PAGE of the final preparation of PolaR., Figure S2: Antibacterial activity of PolaR against Rothia aeria PCM 2669 in time, Figure S3: Antibacterial activity of 150 μg/mL PolaR against Rothia mucilaginosa PCM 2415 (ATCC 25296) was tested over time in a turbidity reduction assay (TRA).

Techniques: Activity Assay, Sonication, Fluorescence

Journal: Biomedicines

Article Title: A Matrix-Free 3D Hepatocyte–Macrophage Co-Culture Spheroid Model for Dual Assessment of Lipid Accumulation and NF-κB-Mediated Inflammatory Activation Under Glucolipotoxic Stress

doi: 10.3390/biomedicines14040792

Figure Lengend Snippet: Morphology and metabolic activity of HepG2/THP-1 spheroids at different seeding densities. ( A ) Representative Live/Dead staining at Day 8 of co-culture spheroids seeded at 3000 (3k) or 6000 (6k) total cells/well. Viable cells are calcein-AM-positive (green) and dead cells are EthD-1-positive (red); corresponding nuclear counterstaining (DAPI, blue) is shown in the lower panels. Scale bars: 100 µm. ( B ) Metabolic activity during maturation (Days 3, 6, 8, and 10) measured by PrestoBlue™ fluorescence (Ex/Em 560/590 nm), normalized to the starting seeded cell number, and reported as normalized RFU. The graph shows one representative independent experiment; data are presented as mean ± SD of 8 technical replicate wells per condition. The experiment was repeated independently three times with comparable results. Two-way ANOVA (factors: time and seeding density) with Šídák’s multiple comparisons. Abbreviations: RFU, relative fluorescence units; EthD-1, ethidium homodimer-1; DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: HepG2 and THP-1 NF-κB-Luc2 were authenticated by STR profiling (ATCC ® Lot number 70050519 and 70059144) and were mycoplasma-negative.

Techniques: Activity Assay, Staining, Co-Culture Assay, Fluorescence

Journal: Biomedicines

Article Title: A Matrix-Free 3D Hepatocyte–Macrophage Co-Culture Spheroid Model for Dual Assessment of Lipid Accumulation and NF-κB-Mediated Inflammatory Activation Under Glucolipotoxic Stress

doi: 10.3390/biomedicines14040792

Figure Lengend Snippet: Hepatocyte and macrophage marker expression during spheroid phenotypic remodeling. HepG2/THP-1 spheroids were harvested at Days 3, 6, 8, and 10 for bulk-spheroid RNA extraction followed by qPCR. ( A ) Hepatocyte-associated transcripts (ALB, AFP, CYP3A4) expressed as fold change versus 2D HepG2 monolayers (calibrator = 1.00). ( B ) Macrophage-associated transcripts (CD14, CD64, CD68, CD206, MARCO, TREM2) expressed as fold change versus 2D PMA-differentiated THP-1 cells (calibrator = 1.00). ( C ) Representative immunofluorescence at Day 8 showing albumin (ALB, green) and CD14 (red); nuclei counterstained with DAPI (blue). Scale bars: 100 µm. The graph shows one representative independent experiment; data are presented as mean ± SD of n = 4 biological replicates per condition, each biological replicate generated by pooling 2 spheroids. The experiment was repeated independently three times with comparable results. One-way ANOVA with Tukey’s multiple comparisons; groups not sharing letters differ significantly ( p < 0.05). Abbreviations: ALB, albumin; AFP, alpha-fetoprotein; CYP3A4, cytochrome P450 family 3 subfamily A member 4; CD, cluster of differentiation; MARCO, macrophage receptor with collagenous structure; TREM2, triggering receptor expressed on myeloid cells 2; DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: HepG2 and THP-1 NF-κB-Luc2 were authenticated by STR profiling (ATCC ® Lot number 70050519 and 70059144) and were mycoplasma-negative.

Techniques: Marker, Expressing, RNA Extraction, Immunofluorescence, Generated