human aortic adventitial fibroblasts haoafs  (PromoCell)

Journal: Rheumatology (Oxford, England)

Article Title: Distinct landscapes of fibroblast subtypes in arteries of patients with giant cell arteritis

doi: 10.1093/rheumatology/keaf143

Figure Lengend Snippet: The expression of fibroblast phenotypic markers in temporal artery biopsies (TAB). ( A ) Representative picture of the immunohistochemical (IHC) staining of CD90, fibroblast activation protein α (FAP), podoplanin (PDPN), CD248 and α-smooth muscle actin (α-SMA) in giant cell arteritis (GCA)-affected TAB. ( B ) IHC staining in control TAB. ( C ) Scoring of fibroblast markers in adventitia, media, and intima (percentage of positive cells per layer). NS, not significant; * P < 0.05, ** P < 0.01. ( D ) Triple immunofluorescence staining of CD90/FAP/PDPN and CD90/α-SMA/CD248 in GCA-affected TAB. PDPN, red; CD90, green; FAP, yellow; CD248, red; α-SMA, yellow; colocalization of CD90 and PDPN is shown in yellow, colocalization of CD90 and FAP is shown in magenta, colocalization of CD90 and αSMA is shown in magenta, colocalization of CD90 and CD248 is shown in cyan. Arrows indicate double positive areas. A, adventitia; M, media; I, intima. For GCA-TAB, intima includes media–intima (MI) and inner intima (II)

Article Snippet: Human aortic adventitia fibroblasts (HAoAF, C-12380) were obtained from PromoCell, Heidelberg, Germany.

Techniques: Expressing, Immunohistochemical staining, Immunohistochemistry, Activation Assay, Control, Immunofluorescence, Staining

Journal: Rheumatology (Oxford, England)

Article Title: Distinct landscapes of fibroblast subtypes in arteries of patients with giant cell arteritis

doi: 10.1093/rheumatology/keaf143

Figure Lengend Snippet: Fibroblast distribution patterns in aorta tissues. ( A ) Expression of fibroblast markers (CD90, fibroblast activation protein [FAP], podoplanin [PDPN], CD248, α-smooth muscle actin [α-SMA]) and cytokines related to remodelling (TGF-β, fibroblast growth factor 21 [FGF21], platelet-derived growth factor B [PDGFB]) in GCA-affected aorta tissues. Adv, adventitia; Med, media; Int, intima. ( B ) Scoring of fibroblast markers and cytokines related to remodelling in adventitia and media. NS, not significant; * P < 0.05, ** P < 0.01. ( C ) Triple immunofluorescence staining of CD90/FAP/PDPN and CD90/α-SMA/CD248 in GCA-affected aorta tissues (structurally disrupted media). CD90, green; PDPN, red; CD24, red; FAP, yellow; α-SMA, yellow. Colocalization of CD90+CD248, CD90+PDPN is shown in cyan; colocalization of CD90+FAP, CD90+α-SMA is shown in magenta. Arrows indicate double positive areas. ( D ) Masson trichrome staining and collagen scoring in aorta tissues. NS: not significant; * P < 0.05

Article Snippet: Human aortic adventitia fibroblasts (HAoAF, C-12380) were obtained from PromoCell, Heidelberg, Germany.

Techniques: Expressing, Activation Assay, Derivative Assay, Immunofluorescence, Staining

Journal: Rheumatology (Oxford, England)

Article Title: Distinct landscapes of fibroblast subtypes in arteries of patients with giant cell arteritis

doi: 10.1093/rheumatology/keaf143

Figure Lengend Snippet: The relationship between fibroblast activating protein (FAP) and human aortic fibroblast proliferation. ( A , B ) Triple immunofluorescent staining of CD90/FAP/Ki67 in GCA-affected temporal artery ( A ) and aorta (structurally disrupted media) ( B ). Ki67, red; CD90, green; FAP, yellow; colocalization of CD90/FAP, cyan; arrows indicate CD90 + FAP + Ki67 + cells. A, adventitia; M, media; I, intima; GCA, giant cell arteritis. ( C ) TGF-β1 5 ng/ml 48 h promoted mRNA expression of FAP and proliferation in human aortic adventitial fibroblasts (HAoAF). Proliferation rate was calculated as Ki67 positive nuclei divided by 4′,6-diamidino-2-phenylindole positive nuclei; Ki67, red. ( D ) HAoAF were transfected with small interfering RNA (siRNA) targeting FAP, and were treated with TGF-β1 6 h later. After 24 h, cells were collected for qPCR; 72 h later, cells were collected for Ki67 staining. FAP knockdown decreased the effect of TGF-β1 on cell proliferation. FAP, red; Ki67, green. NB si: non-binding small interfering RNA; FAP siRNA: small interfering RNA targeting FAP. NS: not significant; * P < 0.05, ** P < 0.01

Article Snippet: Human aortic adventitia fibroblasts (HAoAF, C-12380) were obtained from PromoCell, Heidelberg, Germany.

Techniques: Staining, Expressing, Transfection, Small Interfering RNA, Knockdown, Binding Assay