Journal: Journal of Traditional Chinese Medicine

Article Title: Astragaloside IV plays a role in reducing radiation-induced liver inflammation in mice by inhibiting thioredoxin-interacting protein/nod-like receptor protein 3 signaling pathway

doi: 10.19852/j.cnki.jtcm.2023.01.008

Figure Lengend Snippet: A: immunofluorescence expression of NLRP3 inflammasome in each group. A1: Control group; A2: DMSO group; A3: IR group; A4: IR+AS-IV-20 group; A5: IR+AS-IV-40 group; B1: the expressions of TXNIP, NLRP3, ASC, Caspase-1 and IL-1β in liver were measured by Western blot; B2: relative density of TXNIP; B3: relative density of NLRP3; B4: relative density of ASC. B5: relative density of Caspase-1; B6: relative density of IL-1β. DMSO: dimethyl sulfoxide; IR: irradiation; IR+AS-20: irradiation+AS-IV (20 mg/kg); IR + AS-40: irradiation +AS-IV (40 mg/kg); AS-IV: astragaloside IV; TXNIP: Thioredoxin-interacting protein; NLRP3: nod-like receptor protein 3; ASC: apoptosis-associated speck-like protein containing a CARD; Caspase-1: cysteinyl aspartate-specific proteinase 1; IL-1β: interleukin 1beta. aP < 0.001 vs Control group; bP < 0.001 vs IR group; cP < 0.05 vs IR + AS-20 group; dP < 0.01 vs IR group; eP < 0.01 vs IR + AS-20 group.

Article Snippet: After blocked in 5% nonfat dry milk for 1 h, membranes were washed with Tris-buffered saline with tween 20 (TBST) for 3 × 10 min. Then, membranes were incubated with the primary antibodies against NLRP3 (1:1000), ASC (1:1000; bs-6741R; Bioss, Beijing, China), cysteinyl aspartate-specific proteinase 1 (Caspase-1; 1:1000; bs-10442R; Bioss, Beijing, China), IL-1β (1:1000; bs-0812R; Bioss, Beijing, China), TXNIP (1:1000; ET1705-72, Huabio, Hangzhou, China), tumor necrosis factor alpha (TNF-α, 1:1000; bs-0078R; Bioss, Beijing, China), interleukin 6 (IL-6, 1:1000; bs-0782R; Bioss, Beijing, China)and GAPDH (1:1000; K200057M, Solarbio, Beijing, China) overnight at 4 ℃.

Techniques: Immunofluorescence, Expressing, Western Blot, Irradiation

Journal: Molecular Pain

Article Title: Sodium channel Nav1.7 in vascular myocytes, endothelium, and innervating axons in human skin

doi: 10.1186/s12990-015-0024-3

Figure Lengend Snippet: Nav1.7 immunolabeling (IL) of arterioles (Ar), arteriole-venule shunts (AVS) and associated innervation in normal human plantar glabrous skin with Alomone (A, B) or Yale (C) Nav1.7 antibodies (red). Co-labeling of innervation (arrows) as marked with anti-PGP 9.5 (PGP, green, A ) or smooth muscle cells in tunica media (tm) as marked with anti α-smooth muscle actin antibody (αSMA, green, B , C ). Nuclei are DAPI-labeled (blue). Left images (each panel) show only red fluorescence, middle images green; right images show triple-label combinations. Large white rectangles are 2X-enlargements of small rectangles. A-C . Nav1.7-IL is expressed on endothelial cells of tunica intima (red arrowheads) and tm smooth muscle cells as confirmed by double-labeling with anti-αSMA (B, C) . Nav1.7-IL is expressed on virtually all vascular innervation (arrows) in tunica adventitia (ta) as confirmed by anti-PGP 9.5 double-labeling ( A , yellow arrows). N=nerve. D-E . Nav1.7-IL on arteriole endothelial cells shown as 2X-enlargements of areas indicated by white rectangles in B , C . First images (each panel) show Nav1.7-IL on smooth muscle cells in tm and endothelial cells (red arrowheads). The second images show α-SMA co-labeling of only the smooth muscle cells of tm (green). The third images show merge of first and second images with DAPI (blue). Sections re-labeled with anti-PECAM (green) to show co-labeling with Nav1.7 on endothelial cells (yellow arrowheads, fourth and fifth images). F-G . Background Cy3 fluorescence is limited with no primary antibody in arteriole deep in dermis (F) , epidermis (Ep) and upper-dermis (UD) (G) . In F , broken line shows tm perimeter with dotted line around arteriole lumen. In G , broken line indicates basement membrane of epidermis and dotted line indicates boundary of dead and live superficial keratinocyte layers (stratum corneum, sc and stratum granulosum, sg, respectively). Stratum spinosum, ss; stratum basalis, sb; dermal papilla (dp). Scale bars=150μm (A); 100μm (B ,C, F, G) ; 50μm in D , E .

Article Snippet: Kertinocytes especially in stratum granulosum label for Nav1.7 (arrowheads) which has a more membranous distribution with the Alomone anti-Nav1.7 antibody, but more diffuse labeling with the Abcam anti-Nav1.7 antibody.

Techniques: Immunolabeling, Labeling, Fluorescence

Journal: Molecular Pain

Article Title: Sodium channel Nav1.7 in vascular myocytes, endothelium, and innervating axons in human skin

doi: 10.1186/s12990-015-0024-3

Figure Lengend Snippet: Digital fluorescence images of Nav1.7 immunolabeling (IL) of arterioles (Ar), arteriole-venule shunts (AVS) and associated innervation in normal human glabrous skin biopsies from the plantar foot (A,C,D) and palmar hand (B) . All sections are labeled with an Alomone (A,C) or Yale (B,D) rabbit anti-rat Nav1.7 antibody revealed by a donkey anti-rabbit Cy3-conjugated secondary antibody (red fluorescence). Secondary antibodies conjugated to Alexa 488 (green fluorescence) were used to assess co-labeling for peptidergic sensory innervation revealed with a sheep anti-human CGRP antibody (A,B) or noradrenergic sympathetic innervation revealed with a sheep anti-human NPY antibody (C,D) . Cell nuclei are labeled with DAPI (blue fluorescence). The left images in each panel show only the red fluorescence, the middle images only the green, and the right images the triple label combinations. Areas outlined in large white rectangles are 2X enlargement of the areas in the small rectangles. A-D . Nav1.7-IL is expressed on the endothelial cells of the tunica intima (red arrowheads) and on smooth muscle cells of the tunica media (tm). A , B . Peptidergic sensory innervation co-expresses Nav1.7-IL and CGRP-IL (yellow straight arrows). Other innervation labeled only with Nav1.7 (red curved arrows) is likely the noradrenergic sympathetic innervation that expresses NPY-IL as shown in C and D . C , D . Noradrenergic sympathetic innervation co-expresses Nav1.7-IL and NPY-IL (yellow curved arrows). Other innervation labeled with only Nav1.7 (red straight arrows) is likely the peptidergic sensory innervation that expresses CGRP-IL as shown in A and B . Scale Bar = 150 μm in A and B , 100 μm in C and D .

Article Snippet: Kertinocytes especially in stratum granulosum label for Nav1.7 (arrowheads) which has a more membranous distribution with the Alomone anti-Nav1.7 antibody, but more diffuse labeling with the Abcam anti-Nav1.7 antibody.

Techniques: Fluorescence, Immunolabeling, Labeling

Journal: Molecular Pain

Article Title: Sodium channel Nav1.7 in vascular myocytes, endothelium, and innervating axons in human skin

doi: 10.1186/s12990-015-0024-3

Figure Lengend Snippet: Nav1.7 immunolabeling (IL) of arterioles and associated innervation in normal human palmar glabrous skin biopsies, in alternating sections cut parallel to and through lumen (*) of branched arteriole (A-C) and parallel to arteriole, skimming the interface between tunica media (tm) and tunica adventitia (ta) (D,E) . All sections are labeled with Abcam anti-human Nav1.7 antibody (red). Secondary antibodies conjugated to Alexa 488 (green) were used to assess co-labeling for: smooth muscle cells revealed with mouse anti-α−smooth muscle actin antibody (αSMA, A ); peptidergic sensory innervation revealed with sheep anti-CGRP antibody (yellow straight arrows, B , D ); and noradrenergic sympathetic innervation revealed with sheep anti-NPY antibody (yellow curved arrows, C , E ): Nuclei are labeled with DAPI (blue). Left images in each panel show only the red fluorescence, middle images only green, and right images the triple-label combinations. Areas outlined in large white rectangles (A-C) are 2X enlargements of areas in small rectangles. A-E . A . Nav1.7-IL is expressed on endothelial cells of tunica intima (red arrowheads) and smooth muscle cells of tm as confirmed by double-labeling with anti-αSMA. Nav1.7-IL is expressed on innervation (arrows) in ta, near and at the border with tm. B , D . Peptidergic sensory innervation co-expresses Nav1.7-IL and CGRP-IL (yellow straight arrows). Other innervation labeled only with Nav1.7 (red curved arrows) is likely noradrenergic sympathetic innervation that expresses NPY-IL (C,E) . C , E . Noradrenergic sympathetic innervation co-expresses Nav1.7-IL and NPY-IL (yellow curved arrows). Other innervation labeled with only Nav1.7 (red straight arrows) is likely peptidergic sensory innervation that expresses CGRP-IL as shown in B and D . Scale bar = 100 μm in A-C , 50 μm in D and E .

Article Snippet: Kertinocytes especially in stratum granulosum label for Nav1.7 (arrowheads) which has a more membranous distribution with the Alomone anti-Nav1.7 antibody, but more diffuse labeling with the Abcam anti-Nav1.7 antibody.

Techniques: Immunolabeling, Labeling, Fluorescence

Journal: Molecular Pain

Article Title: Sodium channel Nav1.7 in vascular myocytes, endothelium, and innervating axons in human skin

doi: 10.1186/s12990-015-0024-3

Figure Lengend Snippet: Nav1.7 expression in smooth muscle cells of deep dermis arterioles within skin from lateral malleolus of three healthy subjects. Smooth muscle cells (arrowheads) of the arteriole tunica media exhibit robust Nav1.7 (red) immunolabeling (antibody Nav1.7 Y ), which is co-localized with alpha smooth muscle actin (green). Skin samples from 3 healthy subjects (Subject 1: A ; Subject 2: B , C ; Subject 3: D ) display similar patterns of Nav1.7 labeling in the smooth muscle cells of the dermal arterioles. Co-localization of Nav1.7 and alpha smooth muscle actin is yellow in the merged panels. E . Sections incubated without primary antibodies followed by secondary antibodies displayed background levels of immunofluorescence in skin vasculature.

Article Snippet: Kertinocytes especially in stratum granulosum label for Nav1.7 (arrowheads) which has a more membranous distribution with the Alomone anti-Nav1.7 antibody, but more diffuse labeling with the Abcam anti-Nav1.7 antibody.

Techniques: Expressing, Immunolabeling, Labeling, Incubation, Immunofluorescence

Journal: Molecular Pain

Article Title: Sodium channel Nav1.7 in vascular myocytes, endothelium, and innervating axons in human skin

doi: 10.1186/s12990-015-0024-3

Figure Lengend Snippet: Digital fluorescence images of Nav1.7 (red) and PGP 9.5 (green) immunolabeling (IL) in the epidermis (Ep) and upper dermis (UD) biopsies of normal human palmar glabrous skin ( A , Abcam anti-Nav1.7) and normal human plantar glabrous skin ( B , Alomone anti-Nav1.7). Stratum corneum, sc; stratum granulosum, sg; stratum spinosum (ss); stratum basalis (sb), dermal papilla (dp). Straight arrows indicate epidermal sensory endings, curved arrows indicate small nerves and individual axons or endings in the upper dermis. The areas enclosed in the large rectangles are 2X enlargements of those in the smaller rectangles. Of all the innervation revealed by anti-PGP 9.5, only some express Nav1.7-IL (yellow straight and curved arrows) whereas other only express PGP 9.5-IL (green straight and curved arrows). Aβ-fiber innervation of a Meissner corpuscle (MC) has little if any Nav1.7-IL. Kertinocytes especially in stratum granulosum label for Nav1.7 (arrowheads) which has a more membranous distribution with the Alomone anti-Nav1.7 antibody, but more diffuse labeling with the Abcam anti-Nav1.7 antibody. Scale bar = 100 μm.

Article Snippet: Kertinocytes especially in stratum granulosum label for Nav1.7 (arrowheads) which has a more membranous distribution with the Alomone anti-Nav1.7 antibody, but more diffuse labeling with the Abcam anti-Nav1.7 antibody.

Techniques: Fluorescence, Immunolabeling, Labeling

Journal: Neurobiology of Pain

Article Title: Human-like cutaneous neuropathologies associated with a porcine model of peripheral neuritis: A translational platform for neuropathic pain

doi: 10.1016/j.ynpai.2018.07.002

Figure Lengend Snippet: Representative images of Cy3 immunofluorescent labeling (red) for Nav1.7 (left column), ETA (middle column), and ETB (right column) in two Sham28 (A–F) and two PNT28 (G–L) ipsilateral biopsies. Cell nuclei are counterstained with DAPI (blue fluorescence). E, epidermis; UD, upper dermis; SB, Stratum Basalis; SS, Stratum Spinosum; SG, Stratum Granulosum; SC, Stratum Corneum. Scale bar = 100 μm. A,D,G,J. Nav1.7 immunolableing in Sham28 biopsies was concentrated in SG and upper SS (brackets in A, D) and increased in intensity as well as expanding through the full depth of SS, encroaching on SB in PNT28 biopsies (brackets in G, J). Quantification was based on repeated sampling of average PI in the upper and lower keratinocytes demarcated by the broken and dotted line marquees, respectively. (B, E, H, K) ETA expression in Sham28 biopsies was concentrated in SB (brackets in B, E) and expanded through the full depth of SG and SS in PNT28 biopsies (brackets in H, K). Quantification was based on repeated sampling of average pixel intensities in the SG and SS and in SB demarcated by the broken and dotted line marquees, respectively. (C, F, I, L) ETB expression in Sham28 biopsies was concentrated in SG and upper SS (brackets in C, F) and diminished in PNT28 biopsies (brackets in I, L). Quantification was based on repeated sampling of average pixel intensities in the SG and SS demarcated by the broken line marquees. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Three additional slides from each biopsy were also prepared to detect keratinocyte immunolabeling with specific antibodies against: 2. voltage-gated sodium channel alpha subunit 1.7 ( Nav1.7 ; rabbit polyclonal, 1:500, Alomone).

Techniques: Labeling, Fluorescence, Sampling, Expressing

Journal: Neurobiology of Pain

Article Title: Human-like cutaneous neuropathologies associated with a porcine model of peripheral neuritis: A translational platform for neuropathic pain

doi: 10.1016/j.ynpai.2018.07.002

Figure Lengend Snippet: Keratinocyte average immunofluorescent pixel intensity (PI) from FC18, PC10, PC28, PNT28, and Sham28 for Nav1.7 (A), ETA (C), CGRP (E), and ETB (F). * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.005. (B) Ratio of lower to upper average PI for Nav1.7 (black and gray bars respectively). (D) Ratio of combined SG and SS to SB average PI for ETA (black and gray bars respectively).

Article Snippet: Three additional slides from each biopsy were also prepared to detect keratinocyte immunolabeling with specific antibodies against: 2. voltage-gated sodium channel alpha subunit 1.7 ( Nav1.7 ; rabbit polyclonal, 1:500, Alomone).

Techniques:

Journal: Neurobiology of Pain

Article Title: Human-like cutaneous neuropathologies associated with a porcine model of peripheral neuritis: A translational platform for neuropathic pain

doi: 10.1016/j.ynpai.2018.07.002

Figure Lengend Snippet: (A) Average keratinocyte Nav1.7 PI among upper and lower keratinocytes for ipsilateral biopsies (solid black and gray bars, respectively) and contralateral biopsies (open black and gray bars, respectively) from PNT1, PNT7, PNT14, and PNT 21 pigs (see , Part 2). (B) Ratios of lower divided by upper average keratinocyte Nav1.7 PI for ipsilateral and contralateral biopsies (solid and open bars, respectively). (A) and (B) show results from average ipsilateral Nav1.7 PI among upper and lower epidermal keratinocytes in PNT28 biopsies (solid black and gray bars, respectively) and ipsilateral Sham28 biopsies (open broken line black and gray bars, respectively), included from A and B.

Article Snippet: Three additional slides from each biopsy were also prepared to detect keratinocyte immunolabeling with specific antibodies against: 2. voltage-gated sodium channel alpha subunit 1.7 ( Nav1.7 ; rabbit polyclonal, 1:500, Alomone).

Techniques: