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: Control, Expressing, Staining, Comparison

Journal: PLoS ONE

Article Title: Airway epithelial specific deletion of Jun-N-terminal kinase 1 attenuates pulmonary fibrosis in two independent mouse models

doi: 10.1371/journal.pone.0226904

Figure Lengend Snippet: Primer sequences used in real time PCR analyses.

Article Snippet: Antibodies: Total JNK (#9252), phospho-JNK (#9251) antibodies were obtained from Cell Signaling Technology (Danvers, MA) (all anti Rabbit, 1:1000), antibodies for S100a4 (FSP1) (sc-19949, anti-Goat, 1:500) and actin (sc-8432, anti-mouse 1:5000) were from Santa Cruz Biotechnology (Santa Cruz, CA), antibodies for CCSP (anti goat 1:4000) were a kind gift from Dr. B. Stripp, Cedars-Sinai Medical Center, Los Angeles, CA).

Techniques: Real-time Polymerase Chain Reaction, Amplification

Journal: PLoS ONE

Article Title: Airway epithelial specific deletion of Jun-N-terminal kinase 1 attenuates pulmonary fibrosis in two independent mouse models

doi: 10.1371/journal.pone.0226904

Figure Lengend Snippet: Analysis of mesenchymal ( A ) or epithelial ( B ) mRNA expression in homogenized lung tissue from mice exposed to bleomycin for 3 weeks. Results were normalized to the housekeeping gene cyclophilin, and are expressed as fold expression changes (+/- SEM) compared to the WT vehicle control groups. (WT- Jnk1 : PBS n = 6, Bleo n = 12, ΔEpi Jnk1 : PBS n = 5, Bleo n = 8 respectively mice/group from 2 independent experiments). * p< 0.05 compared to the PBS control group. † p< 0.05 compared to the respective WT group. (ANOVA). C : Evaluation of mesenchymal proteins (α-SMA ( Acta2 ), Col1a1, FSP1 ( S100a4 )) in lungs from ΔEpiJNK1 mice, or control groups subjected to bleomycin-induced lung fibrosis, and the impact of ablation of epithelial JNK1. Homogenized lung tissues were subjected to Western blot analysis for the indicated proteins. β-actin: Loading control. Shown are results from individual mice.

Article Snippet: Antibodies: Total JNK (#9252), phospho-JNK (#9251) antibodies were obtained from Cell Signaling Technology (Danvers, MA) (all anti Rabbit, 1:1000), antibodies for S100a4 (FSP1) (sc-19949, anti-Goat, 1:500) and actin (sc-8432, anti-mouse 1:5000) were from Santa Cruz Biotechnology (Santa Cruz, CA), antibodies for CCSP (anti goat 1:4000) were a kind gift from Dr. B. Stripp, Cedars-Sinai Medical Center, Los Angeles, CA).

Techniques: Expressing, Control, Western Blot

Journal: Mediators of Inflammation

Article Title: Resolution of PMA-Induced Skin Inflammation Involves Interaction of IFN- γ and ALOX15

doi: 10.1155/2013/930124

Figure Lengend Snippet: Neutralization of endogenous IFN- γ decreased the infiltration of inflammatory cells and the production of inflammatory cytokines in the skin. Mice were sacrificed 3 hours, 4 days, or 7 days after the third PMA treatment. (a) Skin sections were stained with anti-CD11b, anti-Gr1, or anti-FSP1 (red). Nucleoli were stained with DAPI (blue). Scale Bar, 50 μ m. (b) The expression levels of IL-6, TNF- α , MCP-1, IL-1 α , and IL-1 β of the skin from control group or anti-IFN- γ group were detected at indicated time points. * P < 0.05, compared to control group.

Article Snippet: For immunofluorescence analysis, sections were stained with rat anti-mouse CD11b (1 : 100; BD Pharmingen, USA), rat anti-mouse granulocyte differentiation antigen 1 (Gr1, 1 : 100; BD Pharmingen, USA), rabbit anti-mouse fibroblast-specific protein-1 (FSP1, 1 : 100; BD Pharmingen, USA), and subsequently rhodamine-labeled secondary antibody and counterstained with 4, 6-diamidino-2-phenylindole (DAPI; Sigma, USA).

Techniques: Neutralization, Staining, Expressing

Journal: Arthritis Research & Therapy

Article Title: Angiotensin II induces skin fibrosis: a novel mouse model of dermal fibrosis

doi: 10.1186/ar4028

Figure Lengend Snippet: Primers for quantitative real-time polymerase chain reaction

Article Snippet: EndoMT , Goat anti-mouse VE-cadherin Ab (Santa Cruz Biotechnology, Santa Cruz, CA); 1:300 , Rabbit anti-mouse FSP1 Ab (Abcam); 1:100 , Alexa fluor 488 donkey anti-goat IgG (Invitrogen); 1:1,000 , Alexa fluor 594 donkey anti-rabbit IgG (Invitrogen); 1:1,000.

Techniques:

Journal: Arthritis Research & Therapy

Article Title: Angiotensin II induces skin fibrosis: a novel mouse model of dermal fibrosis

doi: 10.1186/ar4028

Figure Lengend Snippet: Primary and secondary antibodies for immunofluorescence staining

Article Snippet: EndoMT , Goat anti-mouse VE-cadherin Ab (Santa Cruz Biotechnology, Santa Cruz, CA); 1:300 , Rabbit anti-mouse FSP1 Ab (Abcam); 1:100 , Alexa fluor 488 donkey anti-goat IgG (Invitrogen); 1:1,000 , Alexa fluor 594 donkey anti-rabbit IgG (Invitrogen); 1:1,000.

Techniques: Immunofluorescence

Journal: Arthritis Research & Therapy

Article Title: Angiotensin II induces skin fibrosis: a novel mouse model of dermal fibrosis

doi: 10.1186/ar4028

Figure Lengend Snippet: Recruitment of fibrocytes after angiotensin II treatment . (A) Immunofluorescence staining of CD45 (red) and FSP1 (green) on cryosections of skin samples from PBS- and Ang II-treated mice. Arrows indicate CD45/FSP1 and CD45/P4H double-positive cells. The bar graphs represent quantification of percentage of total positive cells/HPF (high-power field) for CD45, FSP1, CD45/FSP1, and CD45/P4H in PBS- and Ang II-treated mice (* P ≤ 0.05). Representative images are shown from four animals per group. (B) Real-time PCR for MCP1 and FSP1 in PBS (white bars) and Ang II (black bars)-treated mice (* P ≤ 0.05).

Article Snippet: EndoMT , Goat anti-mouse VE-cadherin Ab (Santa Cruz Biotechnology, Santa Cruz, CA); 1:300 , Rabbit anti-mouse FSP1 Ab (Abcam); 1:100 , Alexa fluor 488 donkey anti-goat IgG (Invitrogen); 1:1,000 , Alexa fluor 594 donkey anti-rabbit IgG (Invitrogen); 1:1,000.

Techniques: Immunofluorescence, Staining, Real-time Polymerase Chain Reaction

Journal: Arthritis Research & Therapy

Article Title: Angiotensin II induces skin fibrosis: a novel mouse model of dermal fibrosis

doi: 10.1186/ar4028

Figure Lengend Snippet: Effect of angiotensin II on EndoMT . Immunofluorescence staining of VE-cadherin (green) and FSP1 (red) in skin samples from PBS- and Ang II-treated mice (left panel) and HDMECs treated with 1,000 ng/ml of Ang II and control (right panel). Arrows indicate Ve-cadherin/FSP1 double-positive cells. Representative images are shown from five animals per group.

Article Snippet: EndoMT , Goat anti-mouse VE-cadherin Ab (Santa Cruz Biotechnology, Santa Cruz, CA); 1:300 , Rabbit anti-mouse FSP1 Ab (Abcam); 1:100 , Alexa fluor 488 donkey anti-goat IgG (Invitrogen); 1:1,000 , Alexa fluor 594 donkey anti-rabbit IgG (Invitrogen); 1:1,000.

Techniques: Immunofluorescence, Staining, Control

Journal: CNS neuroscience & therapeutics

Article Title: Oral 7,8-Dihydroxyflavone Protects Retinal Ganglion Cells by Modulating the Gut-Retina Axis and Inhibiting Ferroptosis via the Indoleacrylic Acid-AhR-ALDH1A3-FSP1 Pathway.

doi: 10.1111/cns.70442

Figure Lengend Snippet: FIGURE 5 | IDA inhibits ferroptosis in retinal neurons via the AhR-ALDH1A3-FSP1 pathway. (a) Representative western blot images of f AhR-ALDH1A3-FSP1 pathway proteins. (b–d) Quantification of nAhR, ALDH1A3, and FSP1 levels relative to β-actin. IDA increased nAhR and ALDH1A3 levels, effects partially reversed by the AhR inhibitor (iAhR) but unaffected by ALDH1A3 or FSP1 inhibitors. FSP1 levels remained un- changed. (e–g) Quantification of oxidative stress markers. IDA reduced 4-HNE and MDA levels and increased GSH levels, effects negated by iAhR, iALDH1A3, or iFSP1. Data are mean ± SEM (n = 3). One-way ANOVA with Tukey's post hoc test for (d); unpaired t-test for (b, c, e–g). p values were adjusted for multiple testing by the Benjamini Hochberg method. **p < 0.01, ***p < 0.001 vs. control; ##p < 0.01, ###p < 0.001 vs. IDA group; ns (above bars), p > 0.05 vs. ONC + DHF; ns (above horizontal lines), p > 0.05 among groups below the line.

Article Snippet: IDA's impact on ferroptosis proteins and AhR, ALDH1A3, and FSP1 inhibitors was examined in vitro and in vivo using rabbit anti- AhR (1:1000, #SAB4500725, Sigma), rabbit anti- ALDH1A3 (1:1000, #ABN427, Sigma), rabbit anti- FSP1 (1:1000, #A06541- 2, Boster), rabbit anti- GPX4 (1:1000, #SAB5700944, Sigma), rabbit anti- ACSL4 (1:2000, #SAB2100035, Sigma), rabbit anti- SLC7A11 (1:1000, #SAB5700735, Sigma), rabbit anti- GCH1 (1:1000, #PA5- 103865, Invitrogen), rabbit anti- FTH1 (1:500, #ZRB2695, Sigma), rabbit anti- DHODH (1:1000, #SAB2100574, Sigma), rabbit anti- 4- HNE (1:1000, #MA5- 27570, Invitrogen), and rabbit anti- β- actin (1:2000, #AF5003, Beyotime).

Techniques: Western Blot, Control

Journal: CNS neuroscience & therapeutics

Article Title: Oral 7,8-Dihydroxyflavone Protects Retinal Ganglion Cells by Modulating the Gut-Retina Axis and Inhibiting Ferroptosis via the Indoleacrylic Acid-AhR-ALDH1A3-FSP1 Pathway.

doi: 10.1111/cns.70442

Figure Lengend Snippet: FIGURE 6 | Neuroprotective effects of 7,8-DHF and the gut microbiota-IDA-AhR-ALDH1A3-FSP1 pathway on RGC survival and retinal function following ONC injury. (a) Retinal flat-mount images of RBPMS-labeled RGCs. Scale bar: 100 μm. (b) Representative PhNR traces showing retinal function. (c) Quantification of RGC survival (%). RGC density was reduced in the ONC group compared to the Sham group, partially preserved by 7,8-DHF, attenuated by antibiotics, and restored by IDA. AhR, ALDH1A3, and FSP1 inhibitors partially reduced RGC survival. (d) Quantification of PhNR amplitudes (μV). Retinal function decreased in the ONC group, was partially restored by 7,8-DHF, diminished by antibiotics, and recov- ered by IDA. AhR, ALDH1A3, and FSP1 inhibitors partially reduced the recovery. Data are mean ± SD (n = 6). Unpaired t-test was used except for Sham vs. ONC in (c), where Welch's t-test was applied; +++p < 0.001 vs. Sham; *p < 0.05, ***p < 0.001 vs. ONC; #p < 0.05, ##p < 0.01, ###p < 0.001 vs. ONC + DHF; &&p < 0.01, &&&p < 0.001 vs. ONC + DHF + Abx.

Article Snippet: IDA's impact on ferroptosis proteins and AhR, ALDH1A3, and FSP1 inhibitors was examined in vitro and in vivo using rabbit anti- AhR (1:1000, #SAB4500725, Sigma), rabbit anti- ALDH1A3 (1:1000, #ABN427, Sigma), rabbit anti- FSP1 (1:1000, #A06541- 2, Boster), rabbit anti- GPX4 (1:1000, #SAB5700944, Sigma), rabbit anti- ACSL4 (1:2000, #SAB2100035, Sigma), rabbit anti- SLC7A11 (1:1000, #SAB5700735, Sigma), rabbit anti- GCH1 (1:1000, #PA5- 103865, Invitrogen), rabbit anti- FTH1 (1:500, #ZRB2695, Sigma), rabbit anti- DHODH (1:1000, #SAB2100574, Sigma), rabbit anti- 4- HNE (1:1000, #MA5- 27570, Invitrogen), and rabbit anti- β- actin (1:2000, #AF5003, Beyotime).

Techniques: Labeling

Journal: CNS neuroscience & therapeutics

Article Title: Oral 7,8-Dihydroxyflavone Protects Retinal Ganglion Cells by Modulating the Gut-Retina Axis and Inhibiting Ferroptosis via the Indoleacrylic Acid-AhR-ALDH1A3-FSP1 Pathway.

doi: 10.1111/cns.70442

Figure Lengend Snippet: FIGURE 7 | Effects of 7,8-DHF, gut microbiota disruption, and IDA on AhR-ALDH1A3-FSP1 pathway proteins and oxidative stress in retinal tissues following ONC injury. (a) Representative western blot images for AhR-ALDH1A3-FSP1 pathway proteins. (b–d) Quantification of nAhR, ALDH1A3, and FSP1 levels relative to β-actin. 7,8-DHF increased nAhR and ALDH1A3 levels, which were reduced by antibiotics and restored by IDA. AhR inhibitors decreased nAhR and ALDH1A3, while ALDH1A3 or FSP1 inhibition had no effect on nAhR. FSP1 levels remained unchanged across groups. (e–g) Oxidative stress markers: 7,8-DHF reduced 4-HNE and MDA while increasing GSH levels, effects reversed by antibiotics and restored by IDA. Inhibitors of AhR, ALDH1A3, and FSP1 partially reversed these effects, implicating the AhR-ALDH1A3-FSP1 pathway. Data are mean ± SEM (n = 3). p values were adjusted for multiple testing by the Benjamini Hochberg method. *p < 0.05, **p < 0.01 vs. ONC; #p < 0.05 vs. ONC + DHF; &p < 0.05, &&p < 0.01 vs. ONC + DHF + Abx; ns (above bars), p > 0.05 vs. ONC + DHF; ns (above horizontal lines), p > 0.05 among groups below the line.

Article Snippet: IDA's impact on ferroptosis proteins and AhR, ALDH1A3, and FSP1 inhibitors was examined in vitro and in vivo using rabbit anti- AhR (1:1000, #SAB4500725, Sigma), rabbit anti- ALDH1A3 (1:1000, #ABN427, Sigma), rabbit anti- FSP1 (1:1000, #A06541- 2, Boster), rabbit anti- GPX4 (1:1000, #SAB5700944, Sigma), rabbit anti- ACSL4 (1:2000, #SAB2100035, Sigma), rabbit anti- SLC7A11 (1:1000, #SAB5700735, Sigma), rabbit anti- GCH1 (1:1000, #PA5- 103865, Invitrogen), rabbit anti- FTH1 (1:500, #ZRB2695, Sigma), rabbit anti- DHODH (1:1000, #SAB2100574, Sigma), rabbit anti- 4- HNE (1:1000, #MA5- 27570, Invitrogen), and rabbit anti- β- actin (1:2000, #AF5003, Beyotime).

Techniques: Disruption, Western Blot, Inhibition