Journal: Inflammation and Regeneration

Article Title: Imatinib inhibits pericyte-fibroblast transition and inflammation and promotes axon regeneration by blocking the PDGF-BB/PDGFRβ pathway in spinal cord injury

doi: 10.1186/s41232-022-00223-9

Figure Lengend Snippet: Pericyte transition into fibroblasts after SCI. a – d Representative immunofluorescence images taken in the spinal cords of uninjured mice and injured mice at 3, 7, 14, and 28 days post-injury (dpi) showing that fibrotic scarring PDGFRβ + fibroblasts (red) lose the expression of the pericyte marker NG2 (green, a and b ) but robustly express the fibroblast markers FSP1 (green, c ) and vimentin (green, d ) after SCI. The nuclei are stained with DAPI (blue). The high magnification z-stack images of the dotted area in a are shown below as the region of interest in b. e Quantification of the percentage of NG2 + PDGFRβ + cells out of the total PDGFRβ + cells in the lesion core. f , g Quantification of the percentage of FSP1 + area ( f ) and vimentin + area ( g ). The asterisks indicate the lesion core. Data are shown as mean ± s.e.m. n = 4 mice per time point. Scale bars: 100 μm ( a ) and 10 μm ( b–d ). All images are from sagittal sections. NS, no significance; * p < 0.05, *** p < 0.001 by one-way ANOVA followed by Tukey’s post hoc test in e , f . 3, 7, 14, and 28 dpi versus 0 dpi in e

Article Snippet: The primary antibodies used were as follows: goat anti-CD31(1:100, AF3625, R&D Systems), rat anti-PDGFRβ (1:100, 14-1402-82, Invitrogen), goat anti-PDGFRβ (1:100, AF1042, R&D Systems), rabbit anti-NG2 (1:100, AB5320, Sigma-Aldrich), rabbit anti-FSP1 (1:100, 16105-1-AP, Proteintech), rabbit anti-Vimentin (1:300, ab92547, Abcam), rabbit anti-PDGF-BB (1:50, NBP1-58279, Novus), rat anti-GFAP (1:200, 13-0300, Invitrogen), rabbit anti-GFAP (1:100, 16825-1-AP, Proteintech), rat anti-CD68 (1:300, MCA1957, AbD Serotec), goat anti-5-HT (1:5000, #20080, Immunostar), rabbit anti-neurofilament-heavy polypeptide (NF-H, 1:500, ab207176, Abcam), rabbit anti-growth-associated protein 43 (GAP43, 1:100, 16971-1-AP, Proteintech), rabbit anti-NeuN (1:500, ab177487, Abcam), rabbit anti-fibronectin (1:100, 15613-1-AP, Proteintech), rabbit anti-Laminin (1:100, 23498-1-AP, Proteintech), rat anti-Ki67 (1:100, 14-5698-80, Invitrogen), rabbit anti-cleaved caspase-3 (Asp175) (C-Cas3,1:250, 9661, Cell Signaling Technology), and rabbit anti-fibrinogen (1:100, 15841-12-AP, Proteintech).

Techniques: Immunofluorescence, Expressing, Marker, Staining

Journal: Inflammation and Regeneration

Article Title: Imatinib inhibits pericyte-fibroblast transition and inflammation and promotes axon regeneration by blocking the PDGF-BB/PDGFRβ pathway in spinal cord injury

doi: 10.1186/s41232-022-00223-9

Figure Lengend Snippet: Endothelial PDGF-BB induces pericyte-fibroblast transition and extracellular matrix deposition in vitro. a The PDGF-BB expression levels in bEnd.3 cells treated with 1 mg/ml myelin debris at the indicated time points were measured by ELISA. b , c Western blot analysis ( b ) and quantification ( c ) of PDGF-BB in bEnd.3 cells treated as described above in a . d The PDGF-BB expression levels in bEnd.3 cells treated with the indicated concentrations of myelin debris for 72 h were measured by ELISA. e , f Western blot analysis ( e ) and quantification ( f ) of PDGF-BB in bEnd.3 cells treated as described above in d . g Experimental schematic diagram of pericyte phenotypic transition induced by culture medium (empty, control), EC-CM treated with PBS, Mye-CM treated with myelin debris, and PDGF-BB. h Immunostaining of PDGFRβ (red), pericyte marker NG2 (green, upper panel), and fibroblast markers FSP1 (green, middle panel) and vimentin (green, lower panel) in primary pericytes treated as in g . i Quantification of the percentage of NG2 + , FSP1 + , and vimentin + pericytes. j , k Immunostaining and quantification of extracellular matrix collagen I (green, upper panel) and laminin (green, lower panel) in primary pericytes treated as described in g . The nuclei are stained blue with DAPI. l , m Western blot analysis ( l ) and quantification ( m ) of NG2, FSP1, and vimentin in primary pericytes treated as described above. n , o Western blot analysis ( n ) and quantification ( o ) of extracellular matrix collagen I and laminin in primary pericytes treated as described above. Scale bar: 25 μm ( h and j ). Data are expressed as mean ± s.e.m. n = 3 independent cultures. * p < 0.05, ** p < 0.01, and *** p < 0.001 versus 0 h, 0 mg/ml or control by one-way ANOVA followed by Tukey’s post hoc test in a , c , d , f , i , k , m , and o

Article Snippet: The primary antibodies used were as follows: goat anti-CD31(1:100, AF3625, R&D Systems), rat anti-PDGFRβ (1:100, 14-1402-82, Invitrogen), goat anti-PDGFRβ (1:100, AF1042, R&D Systems), rabbit anti-NG2 (1:100, AB5320, Sigma-Aldrich), rabbit anti-FSP1 (1:100, 16105-1-AP, Proteintech), rabbit anti-Vimentin (1:300, ab92547, Abcam), rabbit anti-PDGF-BB (1:50, NBP1-58279, Novus), rat anti-GFAP (1:200, 13-0300, Invitrogen), rabbit anti-GFAP (1:100, 16825-1-AP, Proteintech), rat anti-CD68 (1:300, MCA1957, AbD Serotec), goat anti-5-HT (1:5000, #20080, Immunostar), rabbit anti-neurofilament-heavy polypeptide (NF-H, 1:500, ab207176, Abcam), rabbit anti-growth-associated protein 43 (GAP43, 1:100, 16971-1-AP, Proteintech), rabbit anti-NeuN (1:500, ab177487, Abcam), rabbit anti-fibronectin (1:100, 15613-1-AP, Proteintech), rabbit anti-Laminin (1:100, 23498-1-AP, Proteintech), rat anti-Ki67 (1:100, 14-5698-80, Invitrogen), rabbit anti-cleaved caspase-3 (Asp175) (C-Cas3,1:250, 9661, Cell Signaling Technology), and rabbit anti-fibrinogen (1:100, 15841-12-AP, Proteintech).

Techniques: In Vitro, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Immunostaining, Marker, Staining

Journal: Inflammation and Regeneration

Article Title: Imatinib inhibits pericyte-fibroblast transition and inflammation and promotes axon regeneration by blocking the PDGF-BB/PDGFRβ pathway in spinal cord injury

doi: 10.1186/s41232-022-00223-9

Figure Lengend Snippet: Microvascular endothelial cells induce pericyte-fibroblast transition via the PDGF-BB/PDGFRβ signaling pathway in vitro. a , b Western blot analysis ( a ) and quantification ( b ) of PDGF-BB in bEnd.3 cells transfected with siNC or siRNAs targeting Pdgfb. c The expression levels of PDGF-BB in bEnd.3 cells transfected with siNC or siPdgfb#2 followed by myelin debris treatment were detected by ELISA. d , e Western blot analysis ( d ) and quantification ( e ) of PDGF-BB in bEnd.3 cells treated as described above in c . f Experimental schematic diagram of pericyte phenotypic transition transfected with siNC or siPdgfb#2 followed by myelin debris treatment. g Immunostaining of NG2 (green, upper panel), FSP1 (green, middle panel), and vimentin (green, lower panel) in primary pericytes treated as described above in f . h Quantification of the percentage of NG2 + , FSP1 + , and vimentin + pericytes in g . i Experimental schematic diagram of pericyte phenotypic transition blocked with the PDGFRβ inhibitor imatinib (a selective PDGFRβ inhibitor) or Su16f (a specific PDGFRβ inhibitor) followed by Mye-CM. j Immunostaining of NG2 (green, upper panel), FSP1 (green, middle panel), and vimentin (green, lower panel) in primary pericytes treated as described in i . k Quantification of the percentage of NG2 + , FSP1 + , and vimentin + pericytes in j . Scale bars: 25 μm ( g and j ). Data are expressed as mean ± s.e.m. n = 3 independent cultures. ** p < 0.01 and *** p < 0.001 by one-way ANOVA followed by Tukey’s post hoc test in b versus siNC, and k . * p < 0.05, ** p < 0.01, and ***p < 0.001 versus siNC by unpaired two-tailed Student’s t test in c , e , and h

Article Snippet: The primary antibodies used were as follows: goat anti-CD31(1:100, AF3625, R&D Systems), rat anti-PDGFRβ (1:100, 14-1402-82, Invitrogen), goat anti-PDGFRβ (1:100, AF1042, R&D Systems), rabbit anti-NG2 (1:100, AB5320, Sigma-Aldrich), rabbit anti-FSP1 (1:100, 16105-1-AP, Proteintech), rabbit anti-Vimentin (1:300, ab92547, Abcam), rabbit anti-PDGF-BB (1:50, NBP1-58279, Novus), rat anti-GFAP (1:200, 13-0300, Invitrogen), rabbit anti-GFAP (1:100, 16825-1-AP, Proteintech), rat anti-CD68 (1:300, MCA1957, AbD Serotec), goat anti-5-HT (1:5000, #20080, Immunostar), rabbit anti-neurofilament-heavy polypeptide (NF-H, 1:500, ab207176, Abcam), rabbit anti-growth-associated protein 43 (GAP43, 1:100, 16971-1-AP, Proteintech), rabbit anti-NeuN (1:500, ab177487, Abcam), rabbit anti-fibronectin (1:100, 15613-1-AP, Proteintech), rabbit anti-Laminin (1:100, 23498-1-AP, Proteintech), rat anti-Ki67 (1:100, 14-5698-80, Invitrogen), rabbit anti-cleaved caspase-3 (Asp175) (C-Cas3,1:250, 9661, Cell Signaling Technology), and rabbit anti-fibrinogen (1:100, 15841-12-AP, Proteintech).

Techniques: In Vitro, Western Blot, Transfection, Expressing, Enzyme-linked Immunosorbent Assay, Immunostaining, Two Tailed Test

Journal: Inflammation and Regeneration

Article Title: Imatinib inhibits pericyte-fibroblast transition and inflammation and promotes axon regeneration by blocking the PDGF-BB/PDGFRβ pathway in spinal cord injury

doi: 10.1186/s41232-022-00223-9

Figure Lengend Snippet: Pharmacologically inhibiting the PDGF-BB/PDGFRβ signaling pathway reduces fibrotic scarring and fibroblasts after SCI. a Representative immunofluorescence images of GFAP (green) and PDGFRβ (red) in mice treated with imatinib or PBS (control) at 14 and 28 days post-injury (dpi). b Representative immunofluorescence images of PDGFRβ (green) and extracellular matrix fibronectin (red, upper panel) and laminin (red, lower panel) at 28 dpi. c , d Quantification of fibrotic scar area ( c ) and extracellular matrix area ( d ) in a and b . e , f Representative immunofluorescence images of PDGFRβ (green) and FSP1 (red) in mice treated as described above at 14 dpi ( e ) and 28 dpi ( f ). The nuclei are stained with DAPI (blue). High magnification images of the dotted area in the left panel are shown in the right panel. All images are from sagittal sections. g Quantification of the fibroblast area occupied by FSP1 in e and f . Scale bars: 100 μm ( a , b and left panel in e and f ) and 20 μm (right panel in e and f ). Data are expressed as mean ± s.e.m. n = 4–6 per group. ** p < 0.01 and *** p < 0.001 versus control by unpaired two-tailed Student’s t test in c , d , and g

Article Snippet: The primary antibodies used were as follows: goat anti-CD31(1:100, AF3625, R&D Systems), rat anti-PDGFRβ (1:100, 14-1402-82, Invitrogen), goat anti-PDGFRβ (1:100, AF1042, R&D Systems), rabbit anti-NG2 (1:100, AB5320, Sigma-Aldrich), rabbit anti-FSP1 (1:100, 16105-1-AP, Proteintech), rabbit anti-Vimentin (1:300, ab92547, Abcam), rabbit anti-PDGF-BB (1:50, NBP1-58279, Novus), rat anti-GFAP (1:200, 13-0300, Invitrogen), rabbit anti-GFAP (1:100, 16825-1-AP, Proteintech), rat anti-CD68 (1:300, MCA1957, AbD Serotec), goat anti-5-HT (1:5000, #20080, Immunostar), rabbit anti-neurofilament-heavy polypeptide (NF-H, 1:500, ab207176, Abcam), rabbit anti-growth-associated protein 43 (GAP43, 1:100, 16971-1-AP, Proteintech), rabbit anti-NeuN (1:500, ab177487, Abcam), rabbit anti-fibronectin (1:100, 15613-1-AP, Proteintech), rabbit anti-Laminin (1:100, 23498-1-AP, Proteintech), rat anti-Ki67 (1:100, 14-5698-80, Invitrogen), rabbit anti-cleaved caspase-3 (Asp175) (C-Cas3,1:250, 9661, Cell Signaling Technology), and rabbit anti-fibrinogen (1:100, 15841-12-AP, Proteintech).

Techniques: Immunofluorescence, Staining, Two Tailed Test

Journal: JCI Insight

Article Title: FGF-2 signaling in nasopharyngeal carcinoma modulates pericyte-macrophage crosstalk and metastasis

doi: 10.1172/jci.insight.157874

Figure Lengend Snippet: ( A ) Cross–data set quantitative heatmap of selected genes of various types of cancer and their adjacent control healthy tissues. Arrow points to distinctively upregulated genes in NPC. Log 2 fold changes were used for quantification. ( B ) Transcriptomic expression levels of FGF2 in human LUAD tissues, BRCA tissues and their adjacent healthy tissues. Sample number: control-LUAD/LUAD/control-BRCA/BRCA=347/483/291/1085. ( C ) Transcriptomic expression levels of FGF2 in various stages of human NPC tissues and their adjacent healthy tissues. Sample number: control/StageT1/StageT2/StageT3=10/16/11/4. ( D ) Human normal nasopharyngeal tissues (NNT), rhinitis tissues, and NPC tissues were stained with H&E and an anti–FGF-2 antibody (brown). Sample number: NNT/Rhinitis/NPC=3/10/6. Scale bar in upper panel: 500 μm. Scale bar in middle and lower panels: 50 μm. Quantification of FGF-2 + signals and FGF-2 + signals in stromal and epithelial components ( n = 8 random fields per group). ( E ) NPC cancer cells were sorted by MACS from freshly tissues. qPCR quantification of FGF2 mRNA ( n = 3 samples per group). ( F ) NNT rhinitis tissues and NPC tissues were stained. Sample number: NNT/Rhinitis/NPC=3/10/6. Scale bar in upper and middle panels: 50 μm. Scale bar in lower panel: 100 μm. Quantification of FSP1 + (brown), CD163 + (brown), CD31 + (red), and NG2 + (green) and coverage rate of NG2 + pericytes ( n = 8 random fields per group). ( G ) qPCR quantification of FGF2 , CD163 , CD31 , NG2 , and FSP1 mRNA in freshly collected tissues. Sample number: Rhinitis/NPC=5/6. ( H ) Correlation of FGF2 and CD163 expression of human NPCs and their control healthy tissues. Sample number: Control/NPC=10/31. * P < 0.05, ** P < 0.01, *** P < 0.001 by unpaired 2-tailed Student’s t test ( B , D , E , G , and H ) or 1-way ANOVA with Tukey’s multiple-comparison analysis ( C , D , and F ). Data are presented as mean ± SD.

Article Snippet: For IHC staining of tumor tissues, paraffin-embedded tissue sections were stained with a rabbit anti–FGF-2 antibody (catalog A0235, ABclonal, 1:100); a mouse anti-FSP1 antibody (catalog 66489-1, Proteintech, 1:100); a rabbit anti-CD163 antibody (catalog A8383, ABclonal, 1:100); a rabbit anti-F4/80 antibody (catalog 70076, Cell Signaling Technology, 1:1000); and a goat anti-CD206 antibody (catalog AF2535, R&D system, 1:400).

Techniques: Expressing, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: A functional role of S100A4/non-muscle myosin IIA axis for pro-tumorigenic vascular functions in glioblastoma

doi: 10.1186/s12964-022-00848-w

Figure Lengend Snippet: Primer sequences of mutant S100A4 promoter used in this study

Article Snippet: Anti-S100A4 (mouse), anti-carbonic anhydrase (CA) 9, and anti-NMIIA antibodies were from Proteintech (Rosemont, IL, USA).

Techniques: Mutagenesis, Sequencing

Journal: Cell Communication and Signaling : CCS

Article Title: A functional role of S100A4/non-muscle myosin IIA axis for pro-tumorigenic vascular functions in glioblastoma

doi: 10.1186/s12964-022-00848-w

Figure Lengend Snippet: Overexpression of S100A4 under hypoxic conditions in GBMs. A Upper: staining by HE and IHC for the indicated proteins in GBMs. Note the strong immunoreactivity for S100A4, NMIIA, HIF-1α, and CA9 in both non-Ps and Ps perinecrotic lesions. Necrotic areas are indicated by asterisks and partitioned by dotted lines. Insets show the magnified views of the boxed area. Original magnification, × 100 and × 400 (inset). Scale bar = 200 μm. Lower: IHC score for the indicated molecules in non-Ps and Ps perinecrotic (non-Ps and Ps) and non-necrotic lesions (non-nec). The data are presented as means ± SDs. B Correlation between IHC scores of S100A4 and related molecules in GBM. ρ , Spearman’s correlation coefficient; n, number of cases

Article Snippet: Anti-S100A4 (mouse), anti-carbonic anhydrase (CA) 9, and anti-NMIIA antibodies were from Proteintech (Rosemont, IL, USA).

Techniques: Over Expression, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: A functional role of S100A4/non-muscle myosin IIA axis for pro-tumorigenic vascular functions in glioblastoma

doi: 10.1186/s12964-022-00848-w

Figure Lengend Snippet: Transcriptional upregulation of S100A4 by HIF-1α. A Western blot (left) and RT-PCR analyses (right) for the indicated molecules in total lysates or total RNA from KS-1 cells treated with 100 and 200 μM CoCl 2 for 24 h. B The S100A4 promoter sequence containing five putative HIF-1α -binding sites (BEs). C KS-1 cells were transfected with Wild type (WT) 1 (− 1986/+ 1012) and WT 2 (− 447/+ 1012) S100A4-luc reporter constructs, together with HIF-1α. Relative activity was determined based on arbitrary luciferase light units normalized to pRL-TK activity. The activities of the reporter plus the effector relative to that of the reporter plus empty vector are shown as means ± SDs. The experiment was performed in duplicate. D Various promoter constructs with mutations (Mut, mutant-type) in the putative HIF-1α-BEs were used for evaluating transcriptional regulation of the S100A4 promoter by HIF-1α. E KS-1 cells were transfected with various mutant constructs of S100A4 promoter, along with HIF-1α

Article Snippet: Anti-S100A4 (mouse), anti-carbonic anhydrase (CA) 9, and anti-NMIIA antibodies were from Proteintech (Rosemont, IL, USA).

Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction, Sequencing, Binding Assay, Transfection, Construct, Activity Assay, Luciferase, Plasmid Preparation, Mutagenesis

Journal: Cell Communication and Signaling : CCS

Article Title: A functional role of S100A4/non-muscle myosin IIA axis for pro-tumorigenic vascular functions in glioblastoma

doi: 10.1186/s12964-022-00848-w

Figure Lengend Snippet: Changes in cell migration ability following knockdown of S100A4 and inhibition of NMIIA activity in GBM cells. A Western blot analysis for the indicated proteins of total lysates from S100A4-knockdown KS-1 (KS-shS100A4#3 and #5) and control cells (Con). B Left: wound-healing assay with S100A4-knockdown KS-1 and control cells (Con). A scratch was made in the middle of confluently grown cells, and phase -contrast images were taken after 0 h (upper), 24 h (middle) and 48 h (lower). Scale bar = 100 μm. Right: the wound areas were calculated using Image J software version 1.41, with the area at 0 h post-wounding set as 1. The fold wound areas are presented as means ± SDs. C Left: migration rate measured using a transwell assay. The S100A4-knockdown KS-1 and control cells (Con) were seeded in a 24-well transwell plate and incubated for 24 h in medium without serum. Cells were stained by HE and counted using a light microscope. Scale bar = 100 μm. Right: cell numbers are presented as means ± SDs (lower). D Left: KS-1 cells were seeded in a 24-well transwell plate and incubated for 24 h in serum-free medium with 0, 2, 4, and 6 μM blebbistatin. Cells were stained by HE and counted using a light microscope. Scale bar = 100 μm. Right: cell numbers are presented as means ± SDs. E Left and middle: in situ PLA assay for the S100A4-NMIIA interaction in non-Ps and Ps perinecrotic lesions of GBMs. Note the small aggregated dots in cytoplasmic and nuclear compartments of the GBM cells in non-Ps, but not Ps, perinecrotic lesions. Necrotic areas are indicated by asterisks and partitioned by dotted lines. Insets show the magnified views of the boxed area. Original magnification, × 100 and × 400 (inset). Scale bar = 200 μm. Right: PLA score for the combinations of S100A4 and NMIIA in non-Ps and Ps perinecrotic (non-Ps and Ps) and non-necrotic lesions (non). The data are presented as means ± SDs

Article Snippet: Anti-S100A4 (mouse), anti-carbonic anhydrase (CA) 9, and anti-NMIIA antibodies were from Proteintech (Rosemont, IL, USA).

Techniques: Migration, Inhibition, Activity Assay, Western Blot, Wound Healing Assay, Software, Transwell Assay, Incubation, Staining, Light Microscopy, In Situ

Journal: Cell Communication and Signaling : CCS

Article Title: A functional role of S100A4/non-muscle myosin IIA axis for pro-tumorigenic vascular functions in glioblastoma

doi: 10.1186/s12964-022-00848-w

Figure Lengend Snippet: Association between S100A4 expression and vascularization in GBM. A Upper: HE staining and IHC for CD34 in non-Ps and Ps perinecrotic and non-necrotic lesions of GBMs. Note the dense CD34 immunoreactivity (closed circle) in non-Ps perinecrotic lesions, in contrast to the scattered immunopositivity (indicated by arrows) in Ps perinecrotic and non-necrotic lesions. Necrotic areas are indicated by asterisks and partitioned by dotted lines. Original magnification, × 100. Scale bar = 200 μm. Lower: microvascular density (MVD) determined by CD34 immunoreactivity in non-Ps and Ps perinecrotic and non-necrotic lesions. The data are presented as means ± SDs. B Left: staining by HE and IHC for the indicate molecules in adjacent, vascular-rich, and distal areas of non-Ps perinecrotic lesions of GBMs. The three lesions are partitioned by dotted lines. Necrotic areas are indicated by asterisks. Insets show the magnified views of the boxed area. Original magnification, × 100 and × 400 (inset). Scale bar = 200 μm. Right: microvascular density (MVD) determined by CD34 immunoreactivity (upper) and IHC score for the indicated molecules in adjacent (adj), vascular-rich (vas), and distal (dis) areas of non-Ps perinecrotic lesions of GBMs. The data are presented as means ± SDs

Article Snippet: Anti-S100A4 (mouse), anti-carbonic anhydrase (CA) 9, and anti-NMIIA antibodies were from Proteintech (Rosemont, IL, USA).

Techniques: Expressing, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: A functional role of S100A4/non-muscle myosin IIA axis for pro-tumorigenic vascular functions in glioblastoma

doi: 10.1186/s12964-022-00848-w

Figure Lengend Snippet: Vascular co-option by S100A4-positive GBM cells and VEGFA mRNA expression in HIF-1α-positive tumor cells. A Staining by HE (upper) and IHC for the indicated proteins (middle and lower) in vascular-rich areas of non-Ps perinecrotic lesions in GBMs. S100A4-positive GBM cells that are co-opted along the surface of preexisting CD34-positive small (indicated by boxes) and mature vessels (indicated by arrows). Original magnification, × 200. Scale bar = 100 μm. B Immunofluorescence for the indicated proteins in vascular-rich area in non-Ps perinecrotic lesion. S100A4/GFAP-positive GBM cells around both small (middle panels: indicated by boxes) and mature vessels (lower panels: indicated by arrows) along SMA-positive vascular components. Original magnification, × 200. Scale bar = 100 μm. C Staining by HE (upper), in situ PLA assay for the S100A4-NMIIA interaction (middle), and IHC for CD34 (lower) in vascular-rich area of a non-Ps perinecrotic lesion in GBM using semiserial sections. Note the small aggregated dots in GBM cells around small vessels (indicated by arrows), as well as those in adjacent areas (indicated by arrow-heads). The three lesions including adjacent, vascular-rich, and distal areas are partitioned by dotted lines. Necrotic areas are indicated by asterisks. Right panels show the magnified views of the boxed areas in the left panels. Original magnification, × 100 (left panels) and × 400 (right panels). Scale bar = 200 μm

Article Snippet: Anti-S100A4 (mouse), anti-carbonic anhydrase (CA) 9, and anti-NMIIA antibodies were from Proteintech (Rosemont, IL, USA).

Techniques: Expressing, Staining, Immunofluorescence, In Situ

Journal: Cell Communication and Signaling : CCS

Article Title: A functional role of S100A4/non-muscle myosin IIA axis for pro-tumorigenic vascular functions in glioblastoma

doi: 10.1186/s12964-022-00848-w

Figure Lengend Snippet: Univariate analyses for overall survival and progression-free survival in GBM

Article Snippet: Anti-S100A4 (mouse), anti-carbonic anhydrase (CA) 9, and anti-NMIIA antibodies were from Proteintech (Rosemont, IL, USA).

Techniques:

Journal: Cell Communication and Signaling : CCS

Article Title: A functional role of S100A4/non-muscle myosin IIA axis for pro-tumorigenic vascular functions in glioblastoma

doi: 10.1186/s12964-022-00848-w

Figure Lengend Snippet: Relationship between S100A4 and NMIIA expression and prognosis in GBMs. A OS (left) and PFS (right) as a function of S100A4 and NMIIA protein expression (upper and lower) in GBM. B TCGA data analyses for OS (left) and PFS (right) as a function of S100A4 and NMIIA mRNA expression (upper and lower) in the GBMs

Article Snippet: Anti-S100A4 (mouse), anti-carbonic anhydrase (CA) 9, and anti-NMIIA antibodies were from Proteintech (Rosemont, IL, USA).

Techniques: Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: A functional role of S100A4/non-muscle myosin IIA axis for pro-tumorigenic vascular functions in glioblastoma

doi: 10.1186/s12964-022-00848-w

Figure Lengend Snippet: Schematic representation of the role of the S100A4/NMIIA axis in GBM. Aggressive and poor prognostic features are elicited through enhancement of tumor cell mobility and acceleration of vascularization

Article Snippet: Anti-S100A4 (mouse), anti-carbonic anhydrase (CA) 9, and anti-NMIIA antibodies were from Proteintech (Rosemont, IL, USA).

Techniques:

Journal: bioRxiv

Article Title: A human fetal lung cell atlas uncovers proximal-distal gradients of differentiation and key regulators of epithelial fates

doi: 10.1101/2022.01.11.474933

Figure Lengend Snippet: (A) UMAP visualization of mesenchymal cells, coloured by cell types. (B) Dot plot describing differential marker gene expression within a cluster by cell type. (C) UMAP visualization of mesenchymal cells, coloured by stages. (D) Visium spatial feature plots visualizing adventitial fibroblasts, vascular SMC 1 and 2 on a 17 pcw lung section. (E-H) In situ HCR assay (E, F, H) and immunostaining (G) for fetal lung tissues at 19 (E), 18 (F), 15 (G), and 17 (H) pcw. (E) Adventitial fibroblasts ( SFRP2 + , white/ PI16 + , red; arrowheads). Endothelial cell ( PECAM1 , green). (F) Alveolar fibroblasts ( WNT2 + , white/ FGFR4 + , red), tip cells ( SFTPC , green). Asterisks (*; myofibroblasts). (G) Airway fibroblasts (S100A4 + , red; dashed lines), tip cells (CD44, green), smooth muscle (ACTA2, white, arrowheads). (H) Myofibroblasts ( KCNK17 + , white/ CXCL14 + , red; arrowheads), tip cells ( SFTPC , green). DAPI, nuclei. Scale bars, 50 µm. (I) UMAP visualization of fibroblast differentiation trajectories, coloured by cell types and stage. (J, K) UMAPs (J) and the relevant gene expression heatmaps (K) displaying potential lineage trajectories from Mid tip to adventitial fibroblasts (top), alveolar fibroblasts (middle), or airway fibroblasts (bottom) derived using Monocle 3.

Article Snippet: After rinsing with cold PBS, treated with primary antibodies against ACTA2 (1:500; Thermo Fisher Scientific, MA1-06110), THBD (1:100; PE-conjugated; BioLegend, 344104), PDGFRA (1:200; Cell Signaling Technology, 3174), S100A4 (1:200; Proteintech, 16105-1-AP), CD44 (1:200; Thermo Fisher Scientific, 17-0441-82), SOX9 (1: 200, Merck, AB5535), PDPN (1:200; R&D Systems, AF3670), or E-cadherin (1: 500; Thermo Fisher Scientific, 13-1900) for 24 hr.

Techniques: Marker, Expressing, In Situ, Host-Cell Reactivation, Immunostaining, Derivative Assay

Journal: bioRxiv

Article Title: A human fetal lung cell atlas uncovers proximal-distal gradients of differentiation and key regulators of epithelial fates

doi: 10.1101/2022.01.11.474933

Figure Lengend Snippet: (A, B) Overview of predicted ligand–receptor interactions from CellPhoneDB in alveolar (A) and airway (B) niches. Dot plots visualize gene expression by cell type and dashed arrows indicate a predicted direction of signaling from ligands to receptors. (C-E) Immunofluorescence of fetal lung tissues at 15 (C) and 19 (D, E) pcw. S100A4/ S100A4 , airway fibroblasts; ACTA2, airway SMCs; CD44, tip epithelium in alveolar region; PECAM1, endothelial cells. S100A4 + airway fibroblasts form a boundary (dashed lines) between alveolar and airway regions. Lines indicate a boundary between airway fibroblasts/airway SMCs and airway epithelial cells. DAPI, nuclei. Scale bars, 50 µm. (F) Experimental outline for airway differentiation of lung tip organoids. Organoids were cultured in FGF7/10-containing medium, in the presence (self-renewal medium; SNM) or absence (differentiation medium; DM) of CHIR99021, a Wnt agonist, for 30 days. (G) Changes in relative mRNA levels of maker genes of the tip progenitors ( SOX9 , SOX2 ), basal cells ( TP63 , KRT5 ), secretory cells ( SCGB3A2 , SCGB1A1 ), and ciliated cells ( FOXJ1 ) by qRT-PCR. Data was normalized to the organoids cultured in SNM. Significance evaluated by 2-way ANOVA with Tukey multiple comparison post-test; * P <0.05, ** P <0.01, *** P <0.001; n = 6 organoid lines. (H) Whole mount immunofluorescence of lung organoids cultured in self-renewal medium (upper) and differentiation medium (lower) for 30 days. DAPI, nuclei. Scale bar, 25 µm.

Article Snippet: After rinsing with cold PBS, treated with primary antibodies against ACTA2 (1:500; Thermo Fisher Scientific, MA1-06110), THBD (1:100; PE-conjugated; BioLegend, 344104), PDGFRA (1:200; Cell Signaling Technology, 3174), S100A4 (1:200; Proteintech, 16105-1-AP), CD44 (1:200; Thermo Fisher Scientific, 17-0441-82), SOX9 (1: 200, Merck, AB5535), PDPN (1:200; R&D Systems, AF3670), or E-cadherin (1: 500; Thermo Fisher Scientific, 13-1900) for 24 hr.

Techniques: Expressing, Immunofluorescence, Cell Culture, Quantitative RT-PCR