Journal: Infection and Immunity

Article Title: Differential sensitivity of leukocyte populations to Staphylococcus aureus biofilm

doi: 10.1128/iai.00654-25

Figure Lengend Snippet: Direct contact with S. aureus biofilm is critical to induce leukocyte cytotoxicity. Primary Mφs, G-MDSCs, and PMNs were co-cultured with S. aureus biofilm either directly or separated by Transwell inserts for the indicated intervals. Leukocytes were stained with anti-CD45, Zombie NIR (viability), Apotracker Green (phosphatidylserine), and MitoSOX Red (mtROS) to quantify ( A ) leukocyte viability, ( B ) early apoptosis, ( C ) late apoptosis/necrosis, and mtROS levels in ( D ) live and ( E ) early apoptotic cells. Unstimulated leukocytes were incubated in medium for 30 min or 2 h, as indicated ( n = 9 from three independent experiments; *, P < 0.05; **, P < 0.01; ****, P < 0.0001; one-way ANOVA with Dunnett’s multiple correction between cell types).

Article Snippet: On day 4 of biofilm growth, ~45% of the medium was removed and leukocytes were added at a density of 2.5 × 10 5 cells/well in fresh medium and incubated for 15 min, 30 min, 2 h, and 6 h. For Transwell co-cultures, Transwell inserts (0.4 μm; CELLTREAT #230635) were placed in 24-well plates above the biofilm on day 4 of growth, whereupon 7.5 × 10 5 leukocytes were added to the upper chamber for 30 min or 2 h. Leukocytes were stained and acquired as described above for planktonic co-culture studies, and unstimulated cells were included for the duration of the co-culture period.

Techniques: Cell Culture, Staining, Incubation

Journal: Nature Communications

Article Title: Purine nucleotide depletion prompts cell migration by stimulating the serine synthesis pathway

doi: 10.1038/s41467-022-30362-z

Figure Lengend Snippet: a Immunoblots of A375 and HeLa cells treated for 24 h with the indicated doses of MTX. b , c Cell-cycle progression through S phase (cell proliferation, BrdU incorporation) ( b ) and relative cell viability (Trypan blue) ( c ) of A375 cells treated with either vehicle (DMSO) or the indicated doses of MTX for 24 h. d Representative images of A375 cells after migration through a transwell (Boyden chamber) treated with either vehicle or the indicated doses of MTX. The migrated cells from five independent biological replicates are presented as relative migration normalized to control (untreated). e Representative images of A375 cells after a wound-healing assay, which were treated with the indicated doses of MTX for 24 h in A375 cells. Quantification of the migrated distance [Gap distance at time 0 h minus gap distance at 24 h] from 6 independent biological replicates. f Representative images of A375 cells after migration through a transwell treated with either vehicle or the indicated doses of GART inhibitor AG2037. The migrated cells were counted from five independent biological replicates and presented as relative migration normalized to control (untreated). g Quantification of the relative migration from a transwell assay in A375 cells treated with vehicle (DMSO) or with the indicated doses of LTX for 24 h as in ( f ). h Representative images from a wound-healing assay of wild-type and ΔGART HeLa cells grown with or without inosine (50 µM) for 24 h. Quantification of the migrated distance was done as in e . i Quantification of the migrated distance from a wound-healing assay (24 h) as in e , in A375 cells treated for 24 h with MTX in the presence or absence of inosine (50 µM). b – i Data are presented as the mean ± s.d from n = 3 ( c ), or n = 5 ( b , d , f , g ) or n = 6 ( e , h , i ) of biologically independent samples. b – i One-way ANOVA, Turkey’s post-hoc test, multiple comparison, ** p < 0.01, *** p < 0.001, **** p < 0.0001. d – f , h Scale bars are indicated. a – i Data are representative of n = 2 ( a ), n = 3 ( b – i ) independent experiments. Source data and exact p -values are provided as a Source Data file.

Article Snippet: Two hundred microliters of cell suspension containing 50,000–120,000 cells were transferred to the Boyden chamber (upper chamber) (CELLTREAT, 230639), while the wells of the 24-well plates (below the chambers) were filled with 500 μl of 10% FBS.

Techniques: Western Blot, BrdU Incorporation Assay, Migration, Control, Wound Healing Assay, Transwell Assay, Comparison

Journal: Pharmaceutics

Article Title: Elastin-like Polypeptide Hydrogels for Tunable, Sustained Local Chemotherapy in Malignant Glioma

doi: 10.3390/pharmaceutics14102072

Figure Lengend Snippet: A qualitative co-localization of DAPI and doxorubicin in U-87 MG cells treated with ELP hydrogels. Cells were plated on poly-lysine treated coverslips placed in wells of a 24-well plate. These were incubated with hydrogels on permeable inserts for 4 h. After treatment coverslips were removed from the wells, washed, and stained with DAPI. A Nikon confocal microscope was used to visualize fluorescence from doxorubicin and DAPI to establish nuclear and perinuclear localization of intracellular doxorubicin.

Article Snippet: The ELP–DOXO hydrogels with three distinct collagen concentrations were made on permeable cell culture inserts (Thincerts, Greiner Bio-one) and allowed to polymerize at 37 °C for 24 h. U-87 MG cells were plated on 24-well format at 10,000 cells per well and incubated overnight to ensure adherence.

Techniques: Incubation, Staining, Microscopy, Fluorescence