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Recruitment and activation of PMNs in UPM-treated mice. ( A ) Hematoxylin and eosin staining and ( B ) immunohistochemistry showed a lot of PMNs ( black arrows ) and CD45-positive cells surrounding the MG acini. ( C ) Immunohistochemistry and ( D ) immunofluorescence of NE disclosed significant upregulation in MGs from the UPM group compared with the control group. ( E – H ) Western blot analysis of ( F ) C5/C5a, ( G ) MMP-9 and ( H ) TNF-α suggested the expression levels of these proteins were all increased (all n = 4 mice per group). Scale bars:, 50 µm. Data are shown as mean ± SD. * p < 0.05 and *** p < 0.001.

Journal: Investigative Ophthalmology & Visual Science

Article Title: Urban Particulate Matter Triggers Meibomian Gland Dysfunction

doi: 10.1167/iovs.65.2.8

Figure Lengend Snippet: Recruitment and activation of PMNs in UPM-treated mice. ( A ) Hematoxylin and eosin staining and ( B ) immunohistochemistry showed a lot of PMNs ( black arrows ) and CD45-positive cells surrounding the MG acini. ( C ) Immunohistochemistry and ( D ) immunofluorescence of NE disclosed significant upregulation in MGs from the UPM group compared with the control group. ( E – H ) Western blot analysis of ( F ) C5/C5a, ( G ) MMP-9 and ( H ) TNF-α suggested the expression levels of these proteins were all increased (all n = 4 mice per group). Scale bars:, 50 µm. Data are shown as mean ± SD. * p < 0.05 and *** p < 0.001.

Article Snippet: The membrane was then blocked in 3% BSA for 2 hours and immunoblotted with primary antibodies overnight: rabbit anti-HMGCR (1:1000, A16875), rabbit anti-SREBP-1 (1:1000, ab3259; Abcam), goat anti-Lrig1 (1:1000, AF3688), rabbit anti-K1 (1:1000, ab185628), rabbit anti-K10 (1:1000, ab76318), rabbit anti-cleaved caspase-8 (1:1000, 8592T; Cell Signaling Technology), rabbit anti-cleaved caspase-9 (1:1000, 20750S; Cell Signaling Technology), rabbit anti-cleaved caspase-3 (1:1000, 9664S), rabbit anti-NLRP3 (1:500, NBP2-12446), rabbit anti-caspase-1 and rabbit anti-cleaved caspase-1 (1:1000, A0964), rabbit anti-GSDMD (1:1000, A18281), rabbit anti-cleaved GSDMD (1:1000, ab209845; Abcam), rabbit anti-IL-18 (1:1000, A1115; Abclonal), rabbit anti-IL-1β (1:1000, ab234437; Abcam), rabbit anti-caspase-4 and rabbit anti-cleaved caspase-4 (1:1000, NBP3-13397; R&D Systems), rabbit anti-C5/C5a (1:1000, A8104; Abclonal), rabbit anti-matrix metalloproteinase-9 (MMP-9) (1:1000, A2095; Abclonal), rabbit anti-TNF-α (1:1000, A0277; Abclonal), rabbit anti-p65 (1:1000, 8242S; Cell Signaling Technology), rabbit anti-phospho-p65 (1:1000, 3033S; Cell Signaling Technology), rabbit anti-p38 (1:1000, 8690S; Cell Signaling Technology), rabbit anti-phospho-p38 (1:1000, 4511S; Cell Signaling Technology), rabbit anti-β-actin (1:2000, 4970S; Cell Signaling Technology), rabbit anti-GAPDH (1:2000, 5174S; Cell Signaling Technology).

Techniques: Activation Assay, Staining, Immunohistochemistry, Immunofluorescence, Western Blot, Expressing