Journal: Investigative Ophthalmology & Visual Science

Article Title: Corneal Nerves Promote Alkali Burn Repair by Modulating Macrophages and Neutrophils via Calcitonin Gene-Related Peptide

doi: 10.1167/iovs.67.4.60

Figure Lengend Snippet: CGRP modulates macrophage function via the cAMP/TSP-1 signaling pathway ( n = 4). (A) Schematic of the cAMP signaling pathway downstream of the CGRP-CALCRL/RAMP1 axis. Activation of adenylyl cyclase (AC) elevates the intracellular level of cAMP. (B) The concentration of cAMP in cell lysates was measured by a competitive ELISA after in vitro stimulation of macrophages with CGRP or co-stimulation of CGRP with SQ22536 for 30 minutes ( n = 4). (C) Bar graph showing quantification of apoptosis in macrophages following in vitro treatment with CGRP alone or in combination with SQ22536 ( n = 4). (D, E) The expression of CD206 and Arg-1 in macrophages after treatment with CGRP or CGRP combined with SQ22536 ( n = 4). (F) Macrophage efferocytosis after CGRP or CGRP + SQ22536 treatment ( n = 4). (G) Volcano plot showing upregulation of thbs1 gene expression in CGRP-treated macrophages. (H, J) Representative micrographs and quantification of increased TSP-1 expression in macrophages following CGRP or CGRP + SQ22536 stimulation in vitro ( n = 4). (I, K) Representative micrographs and quantification of TSP-1+ macrophages in the cornea following 7 days of topical CGRP treatment ( n = 4). (L) Quantification of apoptosis in macrophages treated with TSP-1 in vitro ( n = 4). (M, N) The expression of CD206 and Arg-1 in macrophages after treatment with TSP-1 ( n = 4). (O) Macrophage efferocytosis after CGRP or CGRP + SQ22536 treatment. Data are shown as mean ± SD. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: For TSP-1-induced apoptosis, BMDMs were incubated with RPMI-1640 medium containing 100 nM TSP-1 (HY-P701325; MedChenExpress) for 24 hours.

Techniques: Activation Assay, Concentration Assay, Competitive ELISA, In Vitro, Expressing, Gene Expression

Journal: Drug Design, Development and Therapy

Article Title: Tanshinone IIA Ameliorates Streptozotocin-Induced Diabetic Nephropathy, Partly by Attenuating PERK Pathway-Induced Fibrosis

doi: 10.2147/DDDT.S257734

Figure Lengend Snippet: Tan IIA down-regulates TGF-β1, TSP-1, Grp78 and CHOP expression in the renal tissues of the diabetic rats. ( A ) Immunohistochemical analysis of TGF-β1, TSP-1, Grp78 and CHOP expression in the renal tissues among five groups and the pooled data from ten sections for each group is summarized. Scale bar: 50 μm (×400). ( B, C ) The expression levels of Grp78 ( B ) and CHOP ( C ) from different treatment groups were determined by qRT-PCR. N = 10; * P <0.05 and ** P <0.01.

Article Snippet: The sections were incubated with primary antibodies against Grp78 (1:100), CHOP (1:50), TGF-β1 (1:100) and TSP-1 (1:100) overnight at 4°C, followed by incubating with biotinylated goat anti-rabbit IgG (1:200, # BA1003; Boster Biological Technology, Wuhan, China) in phosphate-buffered saline (PBS) for 2 h at room temperature.

Techniques: Expressing, Immunohistochemical staining, Quantitative RT-PCR