cox2  (Novus Biologicals)

Journal: Scientific reports

Article Title: Tension as a key factor in skin responses to pollution.

doi: 10.1038/s41598-023-42629-6

Figure Lengend Snippet: Figure 6. Ozone exposure upregulates protein levels of COX2 and MMP9, while the presence of tension affects the timing of the response. Ex vivo human skin biopsies were cultured with (tension—Ten) or without (non-tension—NT) tension (Ten Bio models) and then exposed for 4 h to either air or 0.4 ppm of O3. Samples were collected directly after exposure (T0), after 3 h (T3), 6 h (T6) and 24 h (T24). (a) Levels of COX2, green fluorescence staining represents COX2 immunoreactivity. Original magnification ×40. (b) Semi-quantification of the COX2 immunofluorescence intensities performed by ImageJ. (c) Levels of MMP9, red fluorescence staining represents MMP9 immunoreactivity. Original magnification ×40. (d) Semi-quantification of the MMP9 immunofluorescence intensities performed by ImageJ. Results are expressed as arbitrary units ± standard deviation. *p-value < 0.05 by ANOVA.

Article Snippet: Antigen retrieval was achieved via heat-based epitope retrieval with 10 mM sodium citrate buffer (AP-9003-500, Thermo), pH 6.0 with 0.05% Tween 20, at a sub-boiling temperature in a water bath set at 95 °C for 8 min. After cooling for 20 min, the sections were washed 2 X 5 min with 1% Phosphate Buffered Saline (PBS) and then blocked with 5% Bovine Serum Albumin (BSA) in PBS for 45 min. After blocking, sections were then incubated overnight with primary antibodies prepared in 2% BSA/PBS for the following markers: LL-37 (sc-166770, Santa Cruz, dilution 1:50), hBD2 (ab63982, AbCam, dilution 1:500), hBD3 (sc-59495, Santa Cruz, dilution 1:50), 4HNE (AB5605, Millipore, dilution 1:500), AhR (NB 100–128, Novus, 1:150), COX2 (NB 100–868, Novus, 10 μg/ml), MMP9 (NBP2-13173, Novus, 1:200).

Techniques: Ex Vivo, Cell Culture, Fluorescence, Staining, Immunofluorescence, Standard Deviation

Journal: Radiation research

Article Title: Epithelial Responses in Radiation-Induced Lung Injury (RILI) Allow Chronic Inflammation and Fibrogenesis

doi: 10.1667/RADE-22-00103.1

Figure Lengend Snippet: SPC and COX2 expression in lung tissue sections. Panel A: Percent of pro-SPC-expressing cells expressing COX2. Nearly all pro-SPC-positive AEC2 cells express COX2, except within highly cellularized regions detected within 12.5-Gy irradiated animals at 24 weeks. Panel B: Percent of total cells expressing COX2. A significant decrease in the numbers of COX2-positive cells was observed at 16 and 24 weeks in the 12.5-Gy exposure group. In contrast, at 10 Gy, no significant reduction in COX 2 expression as compared to controls was observed. At 24 weeks, COX2 expression was lower within the 12.5-Gy group as compared to the 0- and 10-Gy-exposed animals of similar age. Panel C: Percent of total cells dual labeled for pro-SPC and COX2. A significant decrease in the number of dual positive cells as a percent of total cells was observed at 16 weeks postirradiation after 12.5 Gy, with a further decrease at 24 weeks, which was similarly maintained at 32 weeks. In contrast, no significant difference between the number of dual-labeled cells was observed between the 10-Gy exposure and controls. At 24 and 32 weeks, the number of dual-labeled cells was significantly higher in the 10-Gy group as compared to 12.5 Gy. *P ≤ 0.05 by one-way ANOVA, and Tukey after hoc analysis. Panel D: Representative images showing expression of pro-SPC (green) and COX-2 (red) co-localized (yellow) to DAPI (blue) stained nuclei. Co-localization of pro-SPC, COX2, and DAPI (yellow) expression is reduced in 10- and 12-Gy irradiated animals compared to age matched controls, and noticeably reduced in 12.5 Gy irradiated lungs at 24 and 32 weeks compared to 32-week controls.

Article Snippet: A donkey anti-rabbit secondary antibody (Alexafluor 488; Invitrogen, Eugene, OR) was applied, sections were washed, and again blocked in 5% donkey serum prior to incubation with the second COX2 primary antibody (Novus Biologicals NB100–868, Littleton, CO) in 3% donkey serum in PBS.

Techniques: Expressing, Irradiation, Labeling, Staining

Journal: Radiation research

Article Title: Epithelial Responses in Radiation-Induced Lung Injury (RILI) Allow Chronic Inflammation and Fibrogenesis

doi: 10.1667/RADE-22-00103.1

Figure Lengend Snippet: Transcript abundance in whole lung tissue and CD326+ cells. Panel A: Sftpc transcript (Spc) abundance in whole lung tissue. Spc was significantly reduced at 12, 25, and 32 weeks as compared to 24-h non-irradiated controls. Panel B: Relative Cox2 (Ptgs2) transcript abundance in CD326 cells. Cox2 was significantly decreased at 12 weeks and further reduced in irradiated animals at 24 and 32 weeks. Panel C: Relative Cd200 transcript abundance in CD326 cells. Cd200 was significantly decreased at 4 weeks, and this decrease was maintained at 12 and 16 weeks. *P ≤ 0.05 by one-way ANOVA and pre-selected contrast of means.

Article Snippet: A donkey anti-rabbit secondary antibody (Alexafluor 488; Invitrogen, Eugene, OR) was applied, sections were washed, and again blocked in 5% donkey serum prior to incubation with the second COX2 primary antibody (Novus Biologicals NB100–868, Littleton, CO) in 3% donkey serum in PBS.

Techniques: Irradiation