Journal: Clinical and Vaccine Immunology

Article Title: Increased CXC Ligand 10 Levels and Gene Expression in Type 1 Leprosy Reactions

doi: 10.1128/cvi.00042-11

Figure Lengend Snippet: FIG. 1. Circulating CXCL10 in leprosy patients and healthy con- trols. Each point represents one sample from a different individual. The groups were healthy controls, borderline tuberculoid without re- action (BT), borderline lepromatous without reaction (BL), polar lep- romatous without reaction (LL), BT with reaction (BT plus RR), BL with reaction (BL plus RR), and LL patients with T2R (erythema nodosum leprosum). For patients with reaction (BT plus RR, BL plus RR, and T2R), the sample was taken at the time of reaction, prior to treatment. For LL, n 6; for all other groups, n 10. The horizontal lines indicate median values. The median level of CXCL10 in LL patients with type 2 reaction was not significantly elevated (P 0.9).

Article Snippet: Tissue sections were stained by an indirect immunoperoxidase method using goat anti-human CXCL10 (R&D Systems, Inc.; 1:40) or rabbit anti-human CXCR3 (Sigma) (1:75) for 60 min, followed by biotinylated donkey anti-goat IgG or goat anti-rabbit IgG (both 1:40; Jackson ImmunoResearch Laboratories, Inc.) for 30 min. Peroxidase-conjugated avidin-biotin complexes (Vectastain Elite ABC Reagent kit) were applied for 30 min. All incubations were followed by three washes with phosphate-buffered saline isotonic buffer, pH 7.4.

Techniques:

Journal: Clinical and Vaccine Immunology

Article Title: Increased CXC Ligand 10 Levels and Gene Expression in Type 1 Leprosy Reactions

doi: 10.1128/cvi.00042-11

Figure Lengend Snippet: FIG. 2. Circulating CXCL10 in repeated samples from leprosy patients who developed T1R. Each graph depicts the measurements made from serum samples obtained at 1 month intervals in borderline tuberculoid (A, no. 1 to 10) and borderline lepromatous (B, no. 11 to 20) patients. All patients with clinical T1R received standard multidrug treatment starting at the first visit. The arrows indicate the visits at which T1R was diagnosed. The number above each time point indicates the dose of prednisone started on that day, tapered monthly as indicated; in some cases, corticosteroids were not used initially but were started 1 to 2 months after the initial clinical diagnosis of T1R. Each serum specimen was obtained before corticosteroid treatment was initiated. CXCL10 was significantly elevated during T1R in the 10 BT patients (A) (P 0.0006) and the 10 BL patients (B) (P 0.0001). In patients 12, 16, and 17 (B), T1R was diagnosed by histopathological criteria only; when they were excluded from the analysis, the association of T1R with CXCL10 remained significant (P 0.05).

Article Snippet: Tissue sections were stained by an indirect immunoperoxidase method using goat anti-human CXCL10 (R&D Systems, Inc.; 1:40) or rabbit anti-human CXCR3 (Sigma) (1:75) for 60 min, followed by biotinylated donkey anti-goat IgG or goat anti-rabbit IgG (both 1:40; Jackson ImmunoResearch Laboratories, Inc.) for 30 min. Peroxidase-conjugated avidin-biotin complexes (Vectastain Elite ABC Reagent kit) were applied for 30 min. All incubations were followed by three washes with phosphate-buffered saline isotonic buffer, pH 7.4.

Techniques: Biomarker Discovery

Journal: Clinical and Vaccine Immunology

Article Title: Increased CXC Ligand 10 Levels and Gene Expression in Type 1 Leprosy Reactions

doi: 10.1128/cvi.00042-11

Figure Lengend Snippet: FIG. 3. Expression levels of CXCL10 and IFN- genes in skin biopsy specimens from leprosy patients. (A to H) Real-time quantitative RT-PCR was performed on cDNA obtained from sequential skin biopsy specimens from individual leprosy patients who were placed on MDT, none of whom had T1R at the time of the first biopsy. Five patients (A to E) had clinical symptoms of T1R at the time of the second biopsy; two of these (C and D) had a third biopsy after the reaction had resolved. Three patients (F to H) had no clinical symptoms of T1R at the time of the initial or second biopsy. Data were obtained using the standard-curve method and were normalized using 18S rRNA values, repeated three times for all determinations. The results are presented as the mean standard deviation.

Article Snippet: Tissue sections were stained by an indirect immunoperoxidase method using goat anti-human CXCL10 (R&D Systems, Inc.; 1:40) or rabbit anti-human CXCR3 (Sigma) (1:75) for 60 min, followed by biotinylated donkey anti-goat IgG or goat anti-rabbit IgG (both 1:40; Jackson ImmunoResearch Laboratories, Inc.) for 30 min. Peroxidase-conjugated avidin-biotin complexes (Vectastain Elite ABC Reagent kit) were applied for 30 min. All incubations were followed by three washes with phosphate-buffered saline isotonic buffer, pH 7.4.

Techniques: Expressing, Quantitative RT-PCR, Standard Deviation

Journal: Cell death and differentiation

Article Title: RIG-I-like helicases induce immunogenic cell death of pancreatic cancer cells and sensitize tumors toward killing by CD8(+) T cells.

doi: 10.1038/cdd.2014.96

Figure Lengend Snippet: Figure 1 RLH activation induces secretion of proinflammatory cytokines and induction of apoptosis in murine pancreatic cancer cells. (a) Panc02 cells were stimulated with indicated amounts of ppp-RNA, poly(I:C) or left untreated. OH-RNA served as transfection control. IFN-b levels were analyzed with qRT-PCR relative to HPRT and secretion of CXCL10 or IL-6 was measured with ELISA; (b) Panc02 cells were stimulated with RNA (24 h for poly(I:C) and 48 h for ppp-RNA) and viability was assessed by FACS analysis using annexin V/PI staining; (c) Panc02 cells were incubated with siRNA specific for RIG-I or MDA5 for 24 h and subsequently stimulated with ppp-RNA or poly(I:C). Induction of apoptosis was measured by annexin V/PI staining. Silencing efficacy, as assessed by western blot, is shown; (d) activated caspase-9 (green) was visualized using green FLICA caspase-9 assay kit. Cell membranes were costained with cholera toxin B subunit (red) and nuclei with DAPI (blue); (e and f) Panc02 cells were treated as indicated for 48 h. Full length PARP-1 (116 kDa) and the cleaved large fragment of PARP-1 (89 kDa) (e) as well as the autophagy markers LC3B-I and LC3B-II (f) were analyzed by western blot. Results are representative of at least three independent experiments

Article Snippet: After fixation with paraformaldehyde and permeabilization with saponin (both Sigma), DCs were stained for IL-6 (PE, clone MP5-20F3; BioLegend) or goat anti-mouse CXCL10 (R&D Systems).

Techniques: Activation Assay, Transfection, Control, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Staining, Incubation, Western Blot

Journal: Cell stem cell

Article Title: Chronic inflammation directs an olfactory stem cell functional switch from neuroregeneration to immune defense

doi: 10.1016/j.stem.2019.08.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: The following primary antibodies were used: Rabbit anti-Krt5 (1:800, PRB-160P; Covance), Rabbit anti-Ki67 (1:500, Ab16667; Abcam), Rat anti-BrdU (1:400, Ab6326; Abcam), Mouse anti-Krt14 (1:800, MA5–11599; Thermo Fisher), Mouse anti-β-Tubulin III (1:200, MAB1637; Millipore), Rabbit anti-∆NP63 (1:1,000, 619001; BioLegend), Mouse anti-P63 (1:200, sc-8431; Santa Cruz), Rabbit anti-RelA (1:200, Sc-372; Santa Cruz), Rat anti-CD45 (1:200, 14–0451-81; Ebioscience), Rat anti-F4/80(1:500, MCA497GA; Bio-Rad), Rat anti-Ly6G (1:500, 127601; Biolegend), Rat anti-CD3 (1:200, 14–0032-81; Ebioscience), Goat anti-OMP (1:1000, 544–10001; Wako), Goat anti Sox2 (1:200, sc-17320; Santa Cruz), Goat anti-CCL19 (1:50, AF880; R&D), Goat anti-CXCL10 (1:100, AF-466-NA; R&D), Mouse anti Human CCL2 (1:100, MAB2791; R&D), Rabbit anti Human-CD45 (1:400, Ab40763; Abcam), Mouse anti-CD45 (1:500, 304002; Biolegend), Mouse anti-human CD3 (1:300, 300413; Biolegend), Alexa Fluor® 488 anti-human CD3 (1:200, 300454; Biolegend), Mouse anti-Human CD4 (1: 200, 555344, BD Pharmingen), and Alexa Fluor® 488 anti-β-Tubulin III (1:1000, 801203; Biolegend).

Techniques: Immunohistochemistry, Flow Cytometry, Recombinant, SYBR Green Assay, Enzyme-linked Immunosorbent Assay, RNA Sequencing Assay, Software

Journal: Cellular and Molecular Immunology

Article Title: Synergistic effects of LCN2 and TWEAK on the progression of psoriasis

doi: 10.1038/s41423-025-01292-9

Figure Lengend Snippet: The synergistic interaction between LCN2 and TWEAK in keratinocytes. A Heatmap showing differential expressed genes after co-stimulating with TWEAK and LCN2 in keratinocytes. B Relative expression levels of IL6 , TNFa , IL17A, CCL5, CXCL5, CXCL10 and 24P3R were detected from HFKs using RT-qPCR. C , D Western blot was used to detect p-ERK1/2, p-JNK, p-p38, p-TRAF2, 24P3R and Fn14 protein level in keratinocytes after stimulation with LCN2, TWEAK, or both. One-way ANOVA was used for comparison between groups: ns ( P > 0.05); * P < 0.05; ** P < 0.01; *** P < 0.001

Article Snippet: CXCL10 (1:1000) were bought from GeneTex (GeneTex, Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Comparison

Journal: Cellular and Molecular Immunology

Article Title: Synergistic effects of LCN2 and TWEAK on the progression of psoriasis

doi: 10.1038/s41423-025-01292-9

Figure Lengend Snippet: Lcn 2 –/– mice attenuate the pro-proliferative and inflammatory effects of IMQ on the epidermis. A After being treated with imiquimod and LCN2, the expression levels of Fn14, p-ERK1/2 and in the skin of wild-type mice were analyzed by Western blotting ( n = 3 per group). B Relative expression levels of Tnfsf12(Tweak) , Tnfrsf12a(Fn14) , Il6 , Tnfa , Il1b , Tgfb1 , Krt5 , Krt10 , and Krt17 from mice skin were detected using RT-qPCR. C , D Photographs of skin lesions in mice and the mouse psoriasis severity score difference between wild-type imiquimod mice and Lcn2 –/– imiquimod model mice. Body weight changes in mice modeled with imiquimod for six consecutive days. E Mouse serum levels of TWEAK were assayed by ELISA. F Western blotting was used to detect KRT1, KRT5, KRT6, KRT10, KRT14, KRT16, KRT17, Loricrin, Fn14, CXCL10, LCN2, p-ERK1/2, p-JNK, p-p38 in wild-type and Lcn2 –/– mice after treated with imiquimod ( n = 3 per group). G Relative mRNA expression levels of Lcn2 , Tnfsf12 , Tnfrsf12a , Il6 , Tnfa , Tgfb1 , Krt5 , Krt10 , Krt17 , 24p3r , Mc4r , Il1b were detected by RT-qPCR ( n = 3–5 per group). H Immunostaining of KRT5 and KRT14 were performed on paraffin sections. Data are shown as mean ± SEM ( n = 3–5 per group). ANOVA was used for comparison between groups: ns ( P > 0.05); * P < 0.05; ** P < 0.01; *** P < 0.001

Article Snippet: CXCL10 (1:1000) were bought from GeneTex (GeneTex, Shanghai, China).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Immunostaining, Comparison