Journal: Biomedicines

Article Title: Altered Ocular Surface Health Status and Tear Film Immune Profile Due to Prolonged Daily Mask Wear in Health Care Workers

doi: 10.3390/biomedicines10051160

Figure Lengend Snippet: ( a ) Status of mucin 5AC and mucin 16 in tear fluid in study subjects following mask wear and in corneoscleral explant culture following exposure to hypercapnia. ( b ) Status of mucin 5AC and mucin 16 in corneoscleral explant culture following exposure to hypercapnia.

Article Snippet: The levels of mucins (MUC5AC, MUC16) were measured in the tear fluid of study subjects and the supernatant of the corneoscleral rim explant culture using enzyme-linked immunosorbent assay—ELISA (MUC5AC, E-EL-H2279, Elabscience, Houston, TX, USA; MUC16, E-EL-H0636, Elabscience, Houston, TX, USA) according to the manufacturer’s protocol.

Techniques:

Journal: Biomedicines

Article Title: Altered Ocular Surface Health Status and Tear Film Immune Profile Due to Prolonged Daily Mask Wear in Health Care Workers

doi: 10.3390/biomedicines10051160

Figure Lengend Snippet: Effect of hypercapnia on the expression of mucins in human corneoscleral rim explant cultures. Graphs indicate mean relative mRNA expression of MUC5AC ( a ) and MUC16 ( b ) in matched corneoscleral rims following exposure to either 5% CO 2 (normocapnia—Nor. Cap) or 20% CO 2 (hypercapnia—Hyp. Cap), in vitro for a period of 24 h. The expression of MUC5AC and MUC16 was normalised to expression of β-actin (housekeeping gene). The bar graph indicates mean ± SEM from two technical replicates for each of the three biological replicate experiments. ** p < 0.01; two-tailed paired sample t -test.

Article Snippet: The levels of mucins (MUC5AC, MUC16) were measured in the tear fluid of study subjects and the supernatant of the corneoscleral rim explant culture using enzyme-linked immunosorbent assay—ELISA (MUC5AC, E-EL-H2279, Elabscience, Houston, TX, USA; MUC16, E-EL-H0636, Elabscience, Houston, TX, USA) according to the manufacturer’s protocol.

Techniques: Expressing, In Vitro, Two Tailed Test

Journal: Breast cancer research : BCR

Article Title: Extracellular matrix protein 1 regulates cell proliferation and trastuzumab resistance through activation of epidermal growth factor signaling.

doi: 10.1186/s13058-014-0479-6

Figure Lengend Snippet: Figure 5 Extracellular matrix protein 1 stabilizes epidermal growth factor receptor and HER3 proteins through galectin-3/mucin 1. (A) Lysates from each cell line were analyzed by Western blotting. shC, Control short-hairpin RNA; shE, Extracellular matrix protein 1 short-hairpin RNA; Vec, Vector; WT, Wild type. (B) At 24 hours after seeding, BT-474 trastuzumab-resistant (TR) cells were transfected with each small interfering RNA (siRNA: siC, Control; sigal-3, galectin-3; siMUC1, mucin 1), incubated further for 48 hours and analyzed on Western blots. (C) At 24 hours after seeding, cells were treated with recombinant human extracellular matrix protein 1 (rhECM1; 200 ng/ml) and incubated further for 48 hours. Cell lysates were then incubated with mucin 1 (MUC1), epidermal growth factor receptor (EGFR) and HER3 antibodies overnight. Immunoprecipitates (IP) were analyzed on Western blots. IgG, Immunoglobulin G. (D) Total cell lysates were incubated with MUC1, EGFR and HER3 antibodies overnight, and immunoprecipitates were then analyzed on Western blots.

Article Snippet: ECM1, HER3, phosphorylated ERK (p-ERK), ERK, actin, MUC1 and galectin-3 antibodies were obtained from Santa Cruz Biotechnology. c-Raf, p-c-Raf, MEK, p-MEK, Akt, p-Akt antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Western Blot, Control, shRNA, Plasmid Preparation, Transfection, Small Interfering RNA, Incubation, Recombinant

Journal: Breast cancer research : BCR

Article Title: Extracellular matrix protein 1 regulates cell proliferation and trastuzumab resistance through activation of epidermal growth factor signaling.

doi: 10.1186/s13058-014-0479-6

Figure Lengend Snippet: Figure 6 Extracellular matrix protein 1 induces matrix metalloproteinase 9 transcription. (A) At 24 hours after seeding, cells were incubated with serum-free medium for a further 24 hours. Supernatant medium from each cell line was reacted with matrix metalloproteinase 9 (MMP9) substrate, and relative fluorescence units were determined at 480 to 620 nm. Cont, Control; shC, Control short-hairpin RNA; shE, Extracellular matrix protein 1 short-hairpin RNA; TR, Trastuzumab-resistant; Vec, Vector. (B) MMP9 mRNA levels were determined by real-time PCR using primers specific for MMP9 (*P < 0.05). Each cell line was transfected with an MMP9 promoter luciferase reporter construct. After 48 hours, cells were harvested, and the lysates were analyzed by dual-luciferase assay (*P < 0.05, ***P < 0.0005). (C) At 24 hours after seeding, each cell line was treated with recombinant human MMP9 (rhMMP9; 20 ng/ml), incubated further for 48 hours, and lysates from each cell were analyzed by Western blotting. EGFR, Epidermal growth factor receptor; ERK, Extracellular signal-regulated kinase; MUC1, Mucin 1; WT, Wild type. (D) Schematic model showing the role of extracellular matrix protein 1 (ECM1) in cell signaling. EGF, Epidermal growth factor.

Article Snippet: ECM1, HER3, phosphorylated ERK (p-ERK), ERK, actin, MUC1 and galectin-3 antibodies were obtained from Santa Cruz Biotechnology. c-Raf, p-c-Raf, MEK, p-MEK, Akt, p-Akt antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Incubation, Fluorescence, Control, shRNA, Plasmid Preparation, Real-time Polymerase Chain Reaction, Transfection, Luciferase, Construct, Recombinant, Western Blot