Journal: Clinical & Translational Immunology
Article Title: Naturally derived cytokine peptides limit virus replication and severe disease during influenza A virus infection
doi: 10.1002/cti2.1443
Figure Lengend Snippet: Human growth hormone‐derived LAT8881 improves in vitro cell survival in either a LANCL1 or LANCL2 protein‐dependent manner. (a) Schematic of human growth hormone structure (green, http://www.rcsb.org/structure/1HGU ). The synthetic compound LAT8881 comprises the short C‐terminal region (red) and is cyclised by a disulphide bond between two cysteine residues as shown. (b) Viability of murine L cells (1000 or 5000 cells) relative to vehicle control samples following 24‐ or 48‐h incubation with LAT8881 (200 μM) or vehicle (DMSO) alone, as determined by luminescent ATP detection ± SD. **** P < 0.0001 vs vehicle control, Student's t ‐test. Data are pooled from three independent experiments. (c) Viability of murine peritoneal exudate cells relative to media control in culture at 24 h following LAT8881 (10 μM) or vehicle (DMSO) alone, as determined by luminescent ATP detection ± SD. * P < 0.05, ** P < 0.01 vs media control, two‐way ANOVA. Data are pooled from two independent experiments. (d) Viability of human A549 cells 16 h following 350 μM paclitaxel or vehicle (DMSO) alone ± LAT8881 (1–100 μM), as determined by luminescent ATP detection ± SD. **** P < 0.0001 vs paclitaxel without LAT8881, two‐way ANOVA. Data are representative of two independent experiments. (e) Viability of murine L cells transfected with 100 nM siRNA against Lancl1 or Lancl2 relative to control siRNA samples at 72 or 120 h, as determined by luminescent ATP detection ± SD. * P < 0.05, *** P < 0.001, **** P < 0.0001 vs control siRNA, one‐way ANOVA. Data are pooled from three independent experiments. (f) Viability of human A549 cells transfected with 100 nM siRNA specific to LANCL1 , LANCL2 or control siRNA 16 h following 350 μM paclitaxel ± LAT8881 (1–100 μM) or vehicle, as determined by luminescent ATP detection ± SD. ** P < 0.001, **** P < 0.0001 vs paclitaxel + control siRNA without LAT8881, two‐way ANOVA. Data are representative of two independent experiments.
Article Snippet: In some experiments, L cells were transfected with 100 nM siRNA specific for mouse Lancl1 (Ambion In Vivo , Life Technologies, Carlsbad, USA, Cat #4457308), Lancl2 (Ambion In Vivo , Life Technologies, Carlsbad, USA, Cat #4457308) or control siRNA (Ambion In Vivo Negative Control #1; Cat #4457289) using Lipofectamine 3000 (Thermo Fisher Scientific, Waltham, USA, Cat #L3000015).
Techniques: Derivative Assay, In Vitro, Control, Incubation, Transfection