To visualize the morphology of the C. albicans cells after PDI with PPA-JM-phage, they were examined under scanning electron microscopy (SEM; Hitachi, Tokyo, Japan). C. albicans cells were incubated in 12-well cell culture plates (Sigma Aldrich) with sterile glass coverslips at the bottom for 12 hours at 37°C. Each well was washed three times with sterile PBS and treated with experimental PPA-JM-phage, JM-phage, PPA and control groups (KM13, anti-BSA, PBS). The concentration of PPA in samples was 5 μM, and mixtures were washed three times with PBS to remove unbound phages and PPA. After 1 hour of incubation, C. albicans cells were irradiated with a 658 nm visible red laser under 50 mW/cm2 for 10 minutes. Total power output was 50 mW and spot size 1 cm2. Subsequently, disks were washed three times with PBS and fixed in 2.5% glutaradehyde for 1 hour at 4°C. After fixation, disks were rinsed three times with PBS and dehydrated in an ethanol series (30%, 50%, 70%, 80%, 90% for 7 minutes each, 100% for 10 minutes). Disks were critical point-dried prior to gold sputter-coating and analyzed by SEM.
12 well cell culture plates
Manufactured by Merck Group
The 12-well cell culture plates are a standard laboratory equipment used for culturing cells. They provide a contained and controlled environment for growing and maintaining cells in vitro. Each plate has 12 individual wells, allowing for multiple experiments or conditions to be tested simultaneously. The plates are made of tissue culture-treated polystyrene, which promotes cell attachment and growth. They are designed for use in incubators and other cell culture equipment.
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2 protocols using 12 well cell culture plates
1
SEM Analysis of C. albicans Morphology
2
Oxidative Stress Induction in HaCaT Cells
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HaCaT cells were seeded in 12-well cell culture plates (Sigma-Aldrich) at a density of 1 × 10 5 cells/well in DMEM maintenance medium and incubated for 24 h before cell experiments. The DMEM maintenance medium was replaced with serum-free DMEM supplemented with heat-killed (20 min, 80°C in PBS) bacterial strains at a concentration of 10 8 CFU mL - 1 and incubated for another 24 h for treatment. After 24 h, the medium was replaced with serum-free DMEM supplemented with H 2 O 2 (Sigma-Aldrich) at a valid concentration and time period for inducing oxidative stress as previously described [18] .
Total RNA from the HaCaT cells were isolated using the TRIzol reagent (Thermo Fisher Scienti c) and GeneJET RNA Puri cation Kit (Thermo Fisher Scienti c) following the manufacturer's protocol. Final RNA concentration and quality were determined by ultraviolet absorbance using a NanoDrop spectrophotometer (Thermo Fisher Scienti c). The cDNA was synthesized using a high-capacity cDNA reverse transcription kit (Applied Biosystems). Gene expression levels were quanti ed by qRT-PCR using SYBR® Green (Sigma-Aldrich) and a CFX96 3. Results
Total RNA from the HaCaT cells were isolated using the TRIzol reagent (Thermo Fisher Scienti c) and GeneJET RNA Puri cation Kit (Thermo Fisher Scienti c) following the manufacturer's protocol. Final RNA concentration and quality were determined by ultraviolet absorbance using a NanoDrop spectrophotometer (Thermo Fisher Scienti c). The cDNA was synthesized using a high-capacity cDNA reverse transcription kit (Applied Biosystems). Gene expression levels were quanti ed by qRT-PCR using SYBR® Green (Sigma-Aldrich) and a CFX96 3. Results
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