Journal: bioRxiv

Article Title: Transgene Expression Kinetics and Replication Potential of Recombinant Adenovirus Serotype 4 in a Mouse Model and its Use as a Herpes Simplex Virus Vaccine

doi: 10.64898/2026.05.15.725395

Figure Lengend Snippet: Construction and characterization of Ad4 expressing luciferase or HSV-2 gD. (A) The E3 region of the adenovirus serotype 4 (Ad4) genome is replaced by left and right arms (LA, RA) to allow recombination with a cassette with LA, a CMV promoter, a luciferase or HSV-2 gD gene, a bovine growth hormone (BGH) poly A site, and an RA. The recombinant Ad4 genome is approximately 37 kb in length and retains genes in the E1 region responsible for viral replication. The genome map abbreviations are as follows; left inverted terminal repeat (LITR), E1-early transcriptional gene region 1 (consisting of E1A and E1B), E2-early transcriptional gene region 2 (consisting of E2A and E2B), E3-early transcriptional gene region 3, E4-early transcriptional gene region 4, and right inverted terminal repeat (RITR). (B) In vitro Ad4-Luc infection of A549 cells showing cytopathic effects (MOI 0.1, 3 days after infection, 5x magnification). (C) Uninfected A549 cells for comparison (5x magnification). (D) Relative luminescence of A549 cells infected with Ad4-Luc, Ad4-EGFP, or uninfected. Bars show median values with interquartile range (IQR), with individual data points. ## p=0.009 (Ad4-Luc versus Ad4-EGFP); ** p=0.002 (Ad4-Luc versus uninfected); ns: not significant. (E) Western blot of purified soluble HSV-2 gD protein (Sol gD2), cell lysates of A549 cells infected with Ad4-gD2 (Ad4-gD2), a replicate lysate (Ad4-gD2 rep), and uninfected A549 cells after staining with anti-HSV-2 gD antibody.

Article Snippet: The previously described SpeI site was utilized to linearize pRAd4-EGFP or pRAd4-Luc and this was transfected using PEIpro® (Polyplus Cat# 101000033) into the A549 human carcinoma lung cell line (ATCC cat. CCL-185).

Techniques: Expressing, Luciferase, Recombinant, In Vitro, Infection, Comparison, Western Blot, Purification, Staining

Journal: iScience

Article Title: Drug screening to identify compounds to eliminate Burkholderia pseudomallei through Hcp protein

doi: 10.1016/j.isci.2026.115367

Figure Lengend Snippet: Netilmicin sulfate alone and in combination exerted protective effects on an infected A549 cell model (A) Cytotoxic effects of the drugs on A549 cells. (B) Protective effects of different concentrations of NETS on A549 cells. The early interventions on the abscissa reflect the incubation of A549 cells with the drug for 2 h before B. pseudomallei HNBP001 infection. Late intervention was defined as the introduction of the drug to A549 cells 2 h after infection with B. pseudomallei HNBP001 . The ordinate is the survival rate of the infected A549 cells. NC: the blank negative control group of fresh 10% DMEM. Bp: infected A549 cell group. Bp+30 μM NETS: The infected A549 cells were treated with 30 μM NETS; the other conditions were the same, but different concentrations of NETS were used. (C) Protective effects of several drugs on infected A549 cells; the ordinate is the survival rate of infected A549 cells. NC, blank negative control group of fresh 10% DMEM. Bp: infected A549 cell group. Other: The infected A549 cells were treated with drugs (including SXT, GM, NETS, CAZ, and NETS+CAZ). (D) Number of intracellular bacteria in infected A549 cells after drug treatment; the ordinate represents the number of bacteria in infected A549 cells. NC is the blank negative control group of fresh 10% DMEM. Bp: infected A549 cell group. Drug group: 2 × MIC drug was used to treat infected A549 cells. All drugs, including SXT, GM, NETS, CAZ, and NETS+CAZ. (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001; ns, no significant difference. Data are represented as mean ± SEM.).

Article Snippet: The A549 human lung carcinoma epithelial cell line was purchased from Procell (Wuhan, China; Cat# CL-0016).

Techniques: Infection, Incubation, Negative Control, Bacteria

Journal: iScience

Article Title: Drug screening to identify compounds to eliminate Burkholderia pseudomallei through Hcp protein

doi: 10.1016/j.isci.2026.115367

Figure Lengend Snippet: Netilmicin sulfate alone and in combination exerted protective effects on an infected A549 cell model (A) Cytotoxic effects of the drugs on A549 cells. (B) Protective effects of different concentrations of NETS on A549 cells. The early interventions on the abscissa reflect the incubation of A549 cells with the drug for 2 h before B. pseudomallei HNBP001 infection. Late intervention was defined as the introduction of the drug to A549 cells 2 h after infection with B. pseudomallei HNBP001 . The ordinate is the survival rate of the infected A549 cells. NC: the blank negative control group of fresh 10% DMEM. Bp: infected A549 cell group. Bp+30 μM NETS: The infected A549 cells were treated with 30 μM NETS; the other conditions were the same, but different concentrations of NETS were used. (C) Protective effects of several drugs on infected A549 cells; the ordinate is the survival rate of infected A549 cells. NC, blank negative control group of fresh 10% DMEM. Bp: infected A549 cell group. Other: The infected A549 cells were treated with drugs (including SXT, GM, NETS, CAZ, and NETS+CAZ). (D) Number of intracellular bacteria in infected A549 cells after drug treatment; the ordinate represents the number of bacteria in infected A549 cells. NC is the blank negative control group of fresh 10% DMEM. Bp: infected A549 cell group. Drug group: 2 × MIC drug was used to treat infected A549 cells. All drugs, including SXT, GM, NETS, CAZ, and NETS+CAZ. (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001; ns, no significant difference. Data are represented as mean ± SEM.).

Article Snippet: A549 human lung carcinoma cell line , Procell , Cat#CL-0016.

Techniques: Infection, Incubation, Negative Control, Bacteria

Journal: Materials Today Bio

Article Title: CRISPR-engineered human lung organoids with a biomolecular condensate reporter enable mechanistic toxicity monitoring

doi: 10.1016/j.mtbio.2026.102972

Figure Lengend Snippet: Dose–response profiling of representative toxicant groups in A549 and G3BP1–mCherry hLO. Cell viability was measured after 24 h exposure to five representative toxicants in two in vitro lung models: A549 (2D monolayer) and G3BP1–mCherry knock-in human lung organoids (hLOs). (A) Dose–response curves for the disinfectant/biocide group— didecyldimethylammonium chloride (DDAC), 2-octyl-4-isothiazolin-3-one (OIT), and polyhexamethylene guanidine (PHMG-p). (B) Dose–response curves for industrial chemicals—4,4′-MDI and acrylonitrile. Curves were fitted using nonlinear regression (log[inhibitor] vs . normalized response, variable-slope model) in GraphPad Prism. Data are presented as mean ± SD. (C) Heatmap displaying relative log 2 (IC 50 ) values comparing A549 and hLO models. Each IC 50 value was normalized to the mean IC 50 of the respective compound. The color scale represents deviations from the mean, with blue indicating higher sensitivity and red indicating lower sensitivity. The scale ranges from −2 to +2, corresponding to up to a fourfold deviation from the mean IC 50 ; values outside this range were clipped for clarity.

Article Snippet: A549 human lung epithelial carcinoma cells, purchased from the American Type Culture Collection (Manassas, VA, USA), were cultured in Dulbecco's modified eagle medium, high glucose (LM001-05, WELGENE, Gyeongsan, Korea) supplemented with 10% fetal bovine serum (16000044, Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (15040122, Gibco), 1 mM sodium pyruvate (11360070, Gibco), and 1 × GlutaMAX (35050061, Gibco).

Techniques: In Vitro, Knock-In

Journal: Materials Today Bio

Article Title: CRISPR-engineered human lung organoids with a biomolecular condensate reporter enable mechanistic toxicity monitoring

doi: 10.1016/j.mtbio.2026.102972

Figure Lengend Snippet: Comparative toxicant sensitivity and stress granule responses in G3BP1–mCherry hLOs and conventional lung models. (A) Cell viability analysis of sodium arsenite (NaAsO 2 ), benzalkonium chloride (BAC), and 1,2-benzisothiazolin-3-one (BIT) was evaluated in three lung models: A549 (3D spheroid), patient-derived lung cancer organoid (LCO), and G3BP1–mCherry knock-in lung organoid (hLO). IC 50 values were determined by nonlinear regression (log[inhibitor] vs . normalized response, variable-slope model) using GraphPad Prism. Data are presented as mean ± SD. (B) Heatmap depicting relative log 2 (IC 50 ) values to illustrate model-specific differences in toxicant sensitivity. Each IC 50 value was normalized to the mean IC 50 of the respective compound. Blue and red represent higher and lower sensitivity relative to the mean, respectively. (C) Immunofluorescence staining showing stress granule (SG) formation in patient-derived LCOs and G3BP1–mCherry hLOs after 1 h exposure to sodium arsenite (0, 50, or 100 μM). G3BP1 (green) indicates SG localization, and nuclei were counterstained with Hoechst 33342 (blue). Representative images at intermediate concentrations (12.5, 25, 75 μM) are shown in . Scale bars, 20 μm. (D) Quantification of SG numbers per cell in patient-derived LCOs and hLOs following sodium arsenite treatment (0–100 μM). Bars represent mean ± SD. Statistical analysis was performed using two-way ANOVA followed by Sidak's multiple comparisons test (ns, not significant; *** p < 0.001; **** p < 0.0001).

Article Snippet: A549 human lung epithelial carcinoma cells, purchased from the American Type Culture Collection (Manassas, VA, USA), were cultured in Dulbecco's modified eagle medium, high glucose (LM001-05, WELGENE, Gyeongsan, Korea) supplemented with 10% fetal bovine serum (16000044, Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (15040122, Gibco), 1 mM sodium pyruvate (11360070, Gibco), and 1 × GlutaMAX (35050061, Gibco).

Techniques: Derivative Assay, Knock-In, Immunofluorescence, Staining