Journal: Molecules

Article Title: L-Arginine Enhances Oral Keratinocyte Proliferation under High-Glucose Conditions via Upregulation of CYP1A1 , SKP2 , and SRSF5

doi: 10.3390/molecules28207020

Figure Lengend Snippet: L-Arginine enhances oral keratinocyte proliferation under high-glucose conditions. ( A ) TIGK cells were cultured in 4-chamber slides treated with 500 μM Arg under high-glucose conditions (48 mM). Cell images were taken 24 and 48 h after treatment and the number of cells was counted in each field. ( B ) Representative images of the cultured cells. ( C ) TIGK cells were cultured in 96-well plates. The proliferation was assessed using an MTS proliferation assay. OD490 values were normalized based on the untreated cell condition (100%). Scale bar: 100 µm. Con: Control, Arg: L-Arginine, * p < 0.05, # p < 0.01.

Article Snippet: Cell proliferation was examined using an MTS Cell Proliferation Assay Kit (Abcam, Waltham, MA USA, Catalog # ab197010), according to the manufacturer’s instructions, at 24, 72, and 96 h post-treatment for low-glucose (6 mM) conditions or 72 and 120 h post-treatment for high-glucose (48 mM) conditions.

Techniques: Cell Culture, Proliferation Assay, Control

Journal: Molecules

Article Title: L-Arginine Enhances Oral Keratinocyte Proliferation under High-Glucose Conditions via Upregulation of CYP1A1 , SKP2 , and SRSF5

doi: 10.3390/molecules28207020

Figure Lengend Snippet: Knocking down CYP1A1 , SKP2 , or SRSF5 abolishes enhanced oral keratinocyte proliferation induced by L-Arginine treatment under high-glucose conditions. TIGK cells were transfected with 20 nM siRNAs of CYP1A1 , SKP2 , or SRSF5 and then treated with 500 μM Arg in a culture medium with high glucose (48 mM). ( A ) mRNA expression of CYP1A1 , SKP2 , or SRSF5 48 h after transfection. ( B ) The proliferation of TIGK cells after knocking down CYP1A1 , SKP2 , or SRSF5 assessed by cell count at 48 h and MTS assay at 72 and 120 h after transfection. ( C ) Representative images of cell density at 48 h after treatment. Scale bar: 100 µm. * p < 0.05, # p < 0.01.

Article Snippet: Cell proliferation was examined using an MTS Cell Proliferation Assay Kit (Abcam, Waltham, MA USA, Catalog # ab197010), according to the manufacturer’s instructions, at 24, 72, and 96 h post-treatment for low-glucose (6 mM) conditions or 72 and 120 h post-treatment for high-glucose (48 mM) conditions.

Techniques: Transfection, Expressing, Cell Counting, MTS Assay

Journal: bioRxiv

Article Title: Temperature elevation synergises with and enhances the type-I IFN-mediated restriction of MPXV

doi: 10.1101/2023.09.29.560106

Figure Lengend Snippet: (A) Quantitation of DAPI stained nuclei in mock treated HFt cells incubated at 33, 37, 38.5 or 40 °C for 48 h. (B) MTS cell viability assay of mock treated or puromycin (Puro, 1 μg/ml; positive control) treated HFt cells incubated at 37 or 40 °C for 24 h (as indicated). (C to E) Confluent HFt (human foreskin fibroblast), NOK (human normal oral keratinocyte), RPE (human retinal pigmented epithelial), Vero E6 (green monkey kidney epithelial), or CV-1 (green monkey kidney fibroblast) cell monolayers were infected with (C) HSV-1 (MOI 0.002 PFU/cell), (D) VACV (MOI 0.0005 PFU/cell), or (E) MPXV (MOI 0.001 PFU/cell) for 1 h at 37 °C prior to overlay and incubation at 33, 37, 38.5 or 40 °C. Infected cell monolayers were fixed at 48 h and stained with Coomassie Brilliant blue. (C) Quantitation of HSV-1 plaque counts in HFt cells over the temperature incubation range of analysis. Values were normalised to plaque counts at 37 °C. Means and SD shown; p -values shown, one sample two-tailed t test against a theoretical mean of 1. (D/E) Representative images of VACV or MPXV infected HFT, RPE, NOK, Vero E6 (Meso), and CV-1 cell monolayers stained with Coomassie Brilliant blue at 48 h post-infection (hpi). (A to E) N ≥3 independent biological experiments. Raw values presented in S9 data.

Article Snippet: Cell viability was measured using MTS reagent (Abcam, ab197010), as per the manufacturer’s guidelines, and absorbance measurements taken at OD=490 nm using a PHERAstar plate reader (BMG LABTECH).

Techniques: Quantitation Assay, Staining, Incubation, Viability Assay, Positive Control, Infection, Two Tailed Test