Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: TSLP Interferes with Airway Tolerance by Suppressing the Generation of Antigen-specific Regulatory T cells 1

doi: 10.4049/jimmunol.1002503

Figure Lengend Snippet: A, TSLP suppressed murine iTreg differentiation in the presence of TGF-β1. Naïve CD4+CD62L+ T cells were isolated from spleens and lymph nodes, cultured with 10 μg/ml plate-bound anti-CD3 and 2 μg/ml anti-CD28 in the presence of 3 ng/ml TGF-β1 for 5 days. TSLP was used at 10 ng/ml. Data represent mean ± SEM (n = 3). B, TSLP suppressed human iTreg differentiation in the presence of TGF-β1. Naïve CD4+ T cells were isolated from PBMC and cultured in the same condition as (A). Human T cells showed considerable variability in responding to TSLP. A responder is shown on top, a non-responder on the bottom. The percentage inhibition of iTreg differentiation by TSLP from all 20 donor samples is indicated in the graph on the right. C, Purity of naïve CD4+ T cells isolated from human PBMC. Samples of PBMC and purified T cells from the same donor were stained with Blood Dendritic Cell Enumeration Kit (Miltenyi Biotec) and analyzed by flow cytometry. Data represent mean ± SEM (n = 6). D, Quantitative PCR analysis of the expression of human TSLP receptor TSLPR in activated CD4+ T cells, presented relative to naïve cells. Including of TSLP in culture (+TSLP) further enhanced TSLPR expression. Naïve CD4+ T cells were purified from PBMC and cultured in the same condition as (A). Data represent mean ± SEM (n = 6). *: p < 0.05; **: p < 0.01 One way analysis of variance.

Article Snippet: Since it is reported that human TSLP mainly exerts its effect through DCs and had little effect on CD4 + T cells ( 32 ), we examined DC contamination in our purified human naïve CD4+ T cells by Blood Dendritic Cell Enumeration Kit (Miltenyi Biotec).

Techniques: Isolation, Cell Culture, Inhibition, Purification, Staining, Flow Cytometry, Real-time Polymerase Chain Reaction, Expressing

Journal: PLoS ONE

Article Title: Loss of Grainy Head-Like 1 Is Associated with Disruption of the Epidermal Barrier and Squamous Cell Carcinoma of the Skin

doi: 10.1371/journal.pone.0089247

Figure Lengend Snippet: (A) Blood levels of TSLP in Grhl1 +/+ (gray bar) and Grhl1 −/− mice (blue bar), measured with ELISA kit. (B) Levels of expression of antimicrobial peptides S100A8 and S100A9 in the epidermis of Grhl1 +/+ (gray bar) and Grhl1 −/− mice (blue bar), measured with Q-RT-PCR. (C) Representative skin sections of Grhl1 +/+ (top panel) and Grhl1 −/− mice (bottom panel) stained with toluidine blue to visualize dermal mast cells (purple cells). Scale bars represent 200 µm. (D) Quantification of skin infiltration with mast cells for Grhl1 +/+ (gray bar) and Grhl1 −/− mice (blue bar), estimated as numbers of stained cells per 1 mm 2 area of 10 µm thick skin section (using ImageJ software). (A, B, D) Significance (Student’s t-test, p-value) is shown above bars.

Article Snippet: The blood level of TSLP was measured using enzyme-linked immunosorbent assay (ELISA) kit: Quantikine Mouse TSLP (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions, on groups of seven Grhl1 −/− and five Grhl1 +/+ animals, at the age of about 4 months.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Staining, Software

Journal: Molecules

Article Title: Lignans from Machilus thunbergii as Thymic Stromal Lymphopoietin Inhibitors

doi: 10.3390/molecules26164804

Figure Lengend Snippet: The inhibitory effects of isolated compounds 1 – 3 on thymic stromal lymphopoietin (TSLP)/TSLP receptor (TSLPR) interaction by ELISA.

Article Snippet: For the pSTAT5 assay measuring TSLP inhibitory activity, recombinant human TSLP was purchased from R&D Systems (Minneapolis, MN, USA) and BD Cytofix/Cytoperm and Alexa Fluor 647 mouse anti-STAT5 (pY694) were purchased from BD Biosciences (San Diego, CA, USA).

Techniques: Isolation, Enzyme-linked Immunosorbent Assay, Inhibition, Control

Journal: Molecules

Article Title: Lignans from Machilus thunbergii as Thymic Stromal Lymphopoietin Inhibitors

doi: 10.3390/molecules26164804

Figure Lengend Snippet: Molecular docking results of isolated compounds 1 – 3 on TSLP/TSLPR interaction.

Article Snippet: For the pSTAT5 assay measuring TSLP inhibitory activity, recombinant human TSLP was purchased from R&D Systems (Minneapolis, MN, USA) and BD Cytofix/Cytoperm and Alexa Fluor 647 mouse anti-STAT5 (pY694) were purchased from BD Biosciences (San Diego, CA, USA).

Techniques: Isolation

Journal: Molecules

Article Title: Lignans from Machilus thunbergii as Thymic Stromal Lymphopoietin Inhibitors

doi: 10.3390/molecules26164804

Figure Lengend Snippet: Molecular docking results of isolated compounds 1 – 3 against human TSLP in complex with TSLPR and interleukin-7 receptor α chain (IL-7Rα): ( A ) three-dimensional (3D) docking image of compound 1 ; ( B ) two-dimensional (2D) docking image of compound 1 ; ( C ) 3D docking image of compound 2 ; ( D ) 2D image picture of compound 2 ; ( E ) 3D docking image of compound 3 ; ( F ) 2D docking picture of compound 3 .

Article Snippet: For the pSTAT5 assay measuring TSLP inhibitory activity, recombinant human TSLP was purchased from R&D Systems (Minneapolis, MN, USA) and BD Cytofix/Cytoperm and Alexa Fluor 647 mouse anti-STAT5 (pY694) were purchased from BD Biosciences (San Diego, CA, USA).

Techniques: Isolation

Journal: Biomedical Reports

Article Title: Therapeutic potential of a systemically applied humanized monoclonal antibody targeting Toll‑like receptor 2 in atopic‑dermatitis‑like skin lesions in a mouse model

doi: 10.3892/br.2024.1919

Figure Lengend Snippet: Figure 2. Effect of systemic administration of Toma on DNCB‑induced FLG and TSLP expression in BALB/c mice. (A and B) Paraffin‑embedded tissue sections prepared on day 20 were subjected to immunofluorescence staining with (A) anti‑FLG and (B) anti‑TSLP antibodies with a rhodamine‑red‑X‑conjugated secondary antibody (red). The nuclei were counterstained with Hoechst 33258 (blue). Scale bars, 100 µm. ImageJ software was used to measure fluorescent signals (right graphs). The data are expressed as the mean ± SD. (n=6). *P<0.05 and **P<0.01 by Tukey's multiple comparisons test. Toma; Tomaralimab; DNCB, 2,4‑dinitrochlorobenzene; FLG, filaggrin; TSLP, thymic stromal lymphopoietin; NS, not significant.

Article Snippet: Anti‐thymic stromal lymphopoietin (TSLP) Ab (cat. no. NBP1‐76754) was obtained from Novus Biologicals, LLC and anti‐HMGB1 Ab (cat.

Techniques: Expressing, Immunofluorescence, Staining, Software

Journal: Biomedical Reports

Article Title: Therapeutic potential of a systemically applied humanized monoclonal antibody targeting Toll‑like receptor 2 in atopic‑dermatitis‑like skin lesions in a mouse model

doi: 10.3892/br.2024.1919

Figure Lengend Snippet: Figure 4. Effect of DNCB on the expression of pro‑inflammatory cytokines in HaCaT keratinocytes. (A and B) The effect of DNCB on the expression of pro‑inflammatory cytokines was examined using immunoblot analysis based on different (A) doses and (B) time‑points. The band intensities were measured using ImageJ software. The data are expressed as the mean ± SD (n=3). *P<0.05 and **P<0.01 by Dunnett's multiple comparisons test. DNCB, 2,4‑dinitrochlo‑ robenzene; TSLP, thymic stromal lymphopoietin; IL, interleukin; NS, not significant.

Article Snippet: Anti‐thymic stromal lymphopoietin (TSLP) Ab (cat. no. NBP1‐76754) was obtained from Novus Biologicals, LLC and anti‐HMGB1 Ab (cat.

Techniques: Expressing, Western Blot, Software

Journal: Biomedical Reports

Article Title: Therapeutic potential of a systemically applied humanized monoclonal antibody targeting Toll‑like receptor 2 in atopic‑dermatitis‑like skin lesions in a mouse model

doi: 10.3892/br.2024.1919

Figure Lengend Snippet: Figure 5. Effect of Toma on the inhibition of DNCB‑induced pro‑inflammatory cytokines in HaCaT keratinocytes. (A and B) The effect of Tomaralimab on the inhibition of DNCB‑induced pro‑inflammatory cytokines was examined using (A) reverse transcription‑quantitative PCR and (B) immunoblot analysis. The band intensities were measured using ImageJ software. The data are expressed as the mean ± SD (n=3). *P<0.05 and **P<0.01 by Dunnett's multiple comparisons test. Toma, Tomaralimab; DNCB, 2,4‑dinitrochlorobenzene; TSLP, thymic stromal lymphopoietin; IL, interleukin; NS, not significant.

Article Snippet: Anti‐thymic stromal lymphopoietin (TSLP) Ab (cat. no. NBP1‐76754) was obtained from Novus Biologicals, LLC and anti‐HMGB1 Ab (cat.

Techniques: Inhibition, Western Blot, Software

Journal: Journal of clinical medicine

Article Title: TSLP Exacerbates Septic Inflammation via Murine Double Minute 2 (MDM2) Signaling Pathway.

doi: 10.3390/jcm8091350

Figure Lengend Snippet: Figure 1. TSLP is associated with sepsis. (A) TSLP levels in serum of patients with sepsis were analyzed by ELISA. Normal (Healthy volunteers, n = 20); Sepsis (Patients with sepsis, n = 30). A p value indicates the significant difference between normal and sepsis (B) TSLP levels were analyzed in serum of mice following LPS or E. coli injection by ELISA. (C) TSLP levels were analyzed in peritoneal lavage of mice following LPS or E. coli injection by ELISA. (D) TSLP protein levels were analyzed 12 h after LPS (10 mg/kg) or E. coli (1 × 106 CFU) injection by ELISA. (E) TSLP mRNA expression was analyzed 12 h after LPS (10 mg/kg) or E. coli (1 × 106 CFU) injection by real-time PCR (n = 10/group). A p value indicates the significant difference between PBS and LPS. PBS, phosphate-buffered saline; LPS, lipopolysaccharide; TSLP, thymic stromal lymphopoietin.

Article Snippet: Mice were intravenously injected with recombinant mouse TSLP (R&D Systems, 2 μg mixed with PBS) or PBS as a control (Sigma-Aldrich Co.) by referring to the studies of Piliponsky et al. [10] and Guo et al. [20].

Techniques: Enzyme-linked Immunosorbent Assay, Injection, Expressing, Real-time Polymerase Chain Reaction, Saline

Journal: Journal of clinical medicine

Article Title: TSLP Exacerbates Septic Inflammation via Murine Double Minute 2 (MDM2) Signaling Pathway.

doi: 10.3390/jcm8091350

Figure Lengend Snippet: Figure 2. Systemic inflammatory reaction is blunted by the deficiency of TSLP during sepsis. Mice were given either scramble control or TSLP-specific siRNA + atelocollagen mixture. Each level in (A) serum at 4 h and (B) liver homogenate at 12 h following LPS (10 mg/kg) injection was analyzed by ELISA. Adducts were normalized to total protein in liver homogenate. (C) Each level was analyzed in serum (n = 10/group). A p value indicates the significant difference between PBS and LPS. ** p < 0.05 vs. Con siRNA-received and LPS-injected control mice. PBS, phosphate-buffered saline; LPS, lipopolysaccharide; TSLP, thymic stromal lymphopoietin; Con, control; siRNA, small interfering RNA; VEGF, vascular endothelial growth factor; ICAM-1, intercellular adhesion molecule-1; MIP2, macrophage inflammatory protein 2; AST, aspartate aminotransferase; ALT, alanine aminotransferase, BUN, blood urea nitrogen; CK, creatine kinase.

Article Snippet: Mice were intravenously injected with recombinant mouse TSLP (R&D Systems, 2 μg mixed with PBS) or PBS as a control (Sigma-Aldrich Co.) by referring to the studies of Piliponsky et al. [10] and Guo et al. [20].

Techniques: Control, Injection, Enzyme-linked Immunosorbent Assay, Saline, Small Interfering RNA

Journal: Journal of clinical medicine

Article Title: TSLP Exacerbates Septic Inflammation via Murine Double Minute 2 (MDM2) Signaling Pathway.

doi: 10.3390/jcm8091350

Figure Lengend Snippet: Figure 3. TSLP is produced in macrophages. (A) RAW 264.7 cells were stimulated with LPS or E. coli for 24 h. (A, upper panel) Cell viability was analyzed by an MTT assay. (A, lower panel) NO concentration was measured by the Griess method. (B,C) Cells were stimulated with LPS or E. coli (upper panel) for 24 h for ELISA or (lower panel) 8 h for real-time PCR. Data are representative of three independent experiments (n = 5/group). A p value indicates the significant difference between PBS and LPS. LPS, lipopolysaccharide; TSLP, thymic stromal lymphopoietin; NO, nitric oxide.

Article Snippet: Mice were intravenously injected with recombinant mouse TSLP (R&D Systems, 2 μg mixed with PBS) or PBS as a control (Sigma-Aldrich Co.) by referring to the studies of Piliponsky et al. [10] and Guo et al. [20].

Techniques: Produced, MTT Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction

Journal: Journal of clinical medicine

Article Title: TSLP Exacerbates Septic Inflammation via Murine Double Minute 2 (MDM2) Signaling Pathway.

doi: 10.3390/jcm8091350

Figure Lengend Snippet: Figure 4. TSLP is produced via MDM2 signaling in macrophages. (A,B upper panel) The mRNA and (A,B lower panel) protein expressions of MDM2 were analyzed by real-time PCR and Western blot. (C, upper panel) The mRNA and (C, lower panel) protein expressions of MDM2 in liver were analyzed by real-time PCR and Western blot. (D, left) The production of TSLP 24 h and (D, right) mRNA expression of TSLP 8 h after LPS stimulation were analyzed in RAW 264.7 cells by ELISA and real-time PCR. ** p < 0.05 vs. Con siRNA transfection and LPS stimulation. (E) Cell viability was analyzed by an MTT assay. RAW 264.7 cells were treated with nutlin-3a for 2 h and stimulated with LPS (F, left) for 24 h for ELISA and (F, right) 8 h for real-time PCR. Data are representative of three independent experiments (n = 5/group). # p < 0.05 vs. LPS stimulation. (G) Serum at 4 h and (H) liver homogenate at 12 h following LPS injection from nutlin-3a-treated septic mice were subjected to ELISA (n = 10/group). Adducts were normalized to total protein in liver homogenate. 0.001% DMSO was treated as a vehicle negative control for nutlin-3a in LPS-unstimulated group. A p value indicates the significant difference between PBS and LPS. # p < 0.05 vs. LPS-injected mice. PBS, phosphate-buffered saline; LPS, lipopolysaccharide; TSLP, thymic stromal lymphopoietin; Con, control; siRNA, small interfering RNA; MDM2, murine double minute 2.

Article Snippet: Mice were intravenously injected with recombinant mouse TSLP (R&D Systems, 2 μg mixed with PBS) or PBS as a control (Sigma-Aldrich Co.) by referring to the studies of Piliponsky et al. [10] and Guo et al. [20].

Techniques: Produced, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Transfection, MTT Assay, Injection, Negative Control, Saline, Control, Small Interfering RNA

Journal: Journal of clinical medicine

Article Title: TSLP Exacerbates Septic Inflammation via Murine Double Minute 2 (MDM2) Signaling Pathway.

doi: 10.3390/jcm8091350

Figure Lengend Snippet: Figure 5. TSLP-dependent proinflammatory environment predisposes to the development of sepsis. (A) The mRNA expression of TSLPR and IL-7Rα were analyzed in RAW 264.7 cells by real-time PCR. (B) RAW 264.7 cells were stimulated with LPS (0.1 µg/mL) in the absence or presence of neutralizing anti-TSLP antibody (5 µg/mL) or nonimmune IgG (5 µg/mL) for 24 h. # p < 0.05 vs. LPS stimulation. RAW 264.7 cells were transfected with scramble control siRNA or TSLP-specific siRNA. After LPS stimulation, the mRNA expression levels of (C) TSLP, (D) TSLPR, and IL-7Rα were analyzed in the transfected cells by real-time PCR. (E) The transfected RAW 264.7 cells were stimulated with LPS for 24 h. ** p < 0.05 vs. Con siRNA transfection and LPS stimulation. (F) The peritoneal macrophages isolated from TSLP−/−

Article Snippet: Mice were intravenously injected with recombinant mouse TSLP (R&D Systems, 2 μg mixed with PBS) or PBS as a control (Sigma-Aldrich Co.) by referring to the studies of Piliponsky et al. [10] and Guo et al. [20].

Techniques: Expressing, Real-time Polymerase Chain Reaction, Transfection, Control, Isolation

Journal: Journal of clinical medicine

Article Title: TSLP Exacerbates Septic Inflammation via Murine Double Minute 2 (MDM2) Signaling Pathway.

doi: 10.3390/jcm8091350

Figure Lengend Snippet: Figure 6. Pharmacological inhibition of TSLP by cisplatin protects mice against lethal sepsis. (A) RAW 264.7 cells were treated with cisplatin for 2 h, followed by incubation for 24 h with LPS (0.1 µg/mL). Cell viability was analyzed by MTT assay. RAW 264.7 cells were treated with cisplatin for 2 h, followed by incubation for (B, Left) 24 h with LPS (0.1 µg/mL) for analysis of TSLP production and (B, Right) for 8 h with LPS (0.1 µg/mL) for analysis of TSLP mRNA expression. # p < 0.05 vs. LPS stimulation. (C) RAW 264.7 cells were treated with cisplatin for 2 h, followed by incubation for 8 h with LPS (0.1 µg/mL). MDM2 expression in cells lysates was analyzed by Western blot. Each TSLP level in (D) serum at 4 h and (E) liver homogenate at 12 h following LPS (10 mg/kg) injection was analyzed by ELISA. Adducts were normalized to total protein in liver homogenate. (F) MDM2 expression in liver homogenate at 12 h following LPS (10 mg/kg) injection was analyzed by Western blot. For immunoblots, GAPDH was used as a loading control. (G) BUN, CK, and AST levels were analyzed in the serum at 4 h following LPS (10 mg/kg) injection (n = 10/group). PBS was treated as a vehicle negative control for cisplatin in LPS-unstimulated group. A p value indicates the significant difference between PBS and LPS. # p < 0.05 vs. LPS stimulation. # p < 0.05 vs. LPS-injected mice. (H) Survival curve (%) was monitored in mice (n = 10/group) injected intraperitoneally with LPS (60 mg/kg), E. coli (1 × 108 CFU), or TSLP (2 µg) following intravenous injection of cisplatin (100 µg/kg). LPS, lipopolysaccharide; TSLP, thymic stromal lymphopoietin; MDM2, murine double minute 2; BUN, blood urea nitrogen; CK, creatine kinase; AST, aspartate aminotransferase.

Article Snippet: Mice were intravenously injected with recombinant mouse TSLP (R&D Systems, 2 μg mixed with PBS) or PBS as a control (Sigma-Aldrich Co.) by referring to the studies of Piliponsky et al. [10] and Guo et al. [20].

Techniques: Inhibition, Incubation, MTT Assay, Expressing, Western Blot, Injection, Enzyme-linked Immunosorbent Assay, Control, Negative Control

Journal: Journal of clinical medicine

Article Title: TSLP Exacerbates Septic Inflammation via Murine Double Minute 2 (MDM2) Signaling Pathway.

doi: 10.3390/jcm8091350

Figure Lengend Snippet: Figure 7. Graphical abstract of TSLP function during sepsis. Production of TSLP is induced via the MDM2/NF-κB signaling pathway in E. coli-stimulated macrophages. TSLP affects inflammatory cytokines production and organ dysfunction during sepsis. Cisplatin protects mice against lethal sepsis by inhibiting the series of reactions. TSLP can be induced directly by E. coli or other proinflammatory stimuli. Multiple cell types such as neutrophils, dendritic cells, natural killer cells, and T cells might be involved in sepsis.

Article Snippet: Mice were intravenously injected with recombinant mouse TSLP (R&D Systems, 2 μg mixed with PBS) or PBS as a control (Sigma-Aldrich Co.) by referring to the studies of Piliponsky et al. [10] and Guo et al. [20].

Techniques:

Journal: Oncology letters

Article Title: Human thymic stromal lymphopoietin promotes the proliferation and invasion of cervical cancer cells by downregulating microRNA-132 expression.

doi: 10.3892/ol.2017.7260

Figure Lengend Snippet: Figure 1. TSLP downregulates the expression levels of miR‑132 of HeLa and SiHa cells. Following treatment with differing concentrations of rhTSLP (1, 10 and 100 ng/ml), α‑TSLP or α‑TSLPR (5 µg/ml), or no treatment for 48 h, the mRNA expression levels of miR‑132 in (A) HeLa and (B) SiHa cells were analyzed using reverse transcription‑quantitative polymerase chain reaction analysis. *P<0.05 or **P<0.01 vs. the control. TSLP, thymic stromal lymphopoietin; rhTSLP, recombinant human TSLP; α‑TSLP, anti‑human TSLP neutralizing antibody; α‑TSLPR, anti‑human TSLP receptor neutralizing antibody; miR, microRNA; Ctrl, control.

Article Snippet: Following treatment with a range of concentrations of recombinant human TSLP (rhTSLP; 1, 10, 100 ng/ml; R&D Systems, Inc., Minneapolis, MN, USA), the anti-human TSLP neutralizing antibodies (α-TSLP) or anti-human TSLPR neutralizing antibodies (α-TSLPR) (5 μg/ml; R&D Systems, Inc.) at 37 ̊C in a humidified incubator containing 5% CO2 for 48 h, with 1% PBS used as a control.

Techniques: Expressing, Polymerase Chain Reaction, Control, Recombinant

Journal: Oncology letters

Article Title: Human thymic stromal lymphopoietin promotes the proliferation and invasion of cervical cancer cells by downregulating microRNA-132 expression.

doi: 10.3892/ol.2017.7260

Figure Lengend Snippet: Figure 4. TSLP stimulates the proliferation and invasion of HeLa and SiHa cells via the downregulation of miR‑132. The miR‑NC and miR‑132‑mimic HeLa and SiHa cells were treated with or without rhTSLP (10 ng/ml), and then the proliferation and invasion of these cells were analyzed using (A) a BrdU cell proliferation assay for 72 h and (B) a Matrigel invasion assay for 48 h. *P<0.05, **P<0.01 and ***P<0.001 vs. miR‑NC. NS, #P<0.05 or ##P<0.01 vs. the no rhTSLP group. TSLP, thymic stromal lymphopoietin; miR, microRNA; NC, negative control; rhTSLP, recombinant human TSLP; NS, not significant; ctrl, control.

Article Snippet: Following treatment with a range of concentrations of recombinant human TSLP (rhTSLP; 1, 10, 100 ng/ml; R&D Systems, Inc., Minneapolis, MN, USA), the anti-human TSLP neutralizing antibodies (α-TSLP) or anti-human TSLPR neutralizing antibodies (α-TSLPR) (5 μg/ml; R&D Systems, Inc.) at 37 ̊C in a humidified incubator containing 5% CO2 for 48 h, with 1% PBS used as a control.

Techniques: BrdU Cell Proliferation Assay, Invasion Assay, Negative Control, Recombinant, Control

Journal: Oncology letters

Article Title: Human thymic stromal lymphopoietin promotes the proliferation and invasion of cervical cancer cells by downregulating microRNA-132 expression.

doi: 10.3892/ol.2017.7260

Figure Lengend Snippet: Figure 5. Function of TSLP/miR‑132 signaling in CC cells. CC cells secrete high levels of TSLP. TSLP then downregulates the level of miR‑132 in CC cells. As a result, miR‑132 suppresses the expression of proliferation‑associ ated molecules Ki‑67 and PCNA, and invasion‑associated molecules MMP2 and MMP9, and further inhibits the proliferation and invasion of CC cells. Therefore, TSLP/miR‑132 signaling contributes to the progression of CC. TSLP, thymic stromal lymphopoietin; miR, microRNA; CC, cervial cancer; Ki‑67, marker of proliferation Ki‑67; PCNA, proliferating cell nuclear antigen; MMP, matrix metalloproteinase.

Article Snippet: Following treatment with a range of concentrations of recombinant human TSLP (rhTSLP; 1, 10, 100 ng/ml; R&D Systems, Inc., Minneapolis, MN, USA), the anti-human TSLP neutralizing antibodies (α-TSLP) or anti-human TSLPR neutralizing antibodies (α-TSLPR) (5 μg/ml; R&D Systems, Inc.) at 37 ̊C in a humidified incubator containing 5% CO2 for 48 h, with 1% PBS used as a control.

Techniques: Expressing, Marker

Journal: International Journal of Molecular Sciences

Article Title: Blockade of PAR2 Signaling by Punicalagin as a Therapeutic Strategy for Atopic Dermatitis

doi: 10.3390/ijms26188920

Figure Lengend Snippet: PCG suppresses PAR2-induced inflammatory signaling in HaCaT cells. ( A ) Representative immunoblot showing the effect of PCG on ERK1/2 and p65 phosphorylation. HaCaT cells were pretreated with various concentrations of PCG for 20 min before stimulation with 30 µM PAR2-AP. ( B , C ) Quantification of phosphorylated ERK1/2 and p65 levels. Phosphorylated ERK1/2 bands were normalized to total ERK1/2 and phosphorylated p65 (p-p65) was normalized to total p65. ( D ) Effect of PCG on PAR2-AP-induced IL-8 secretion. HaCaT cells were pretreated with PCG for 0 min and then stimulated with 30 µM PAR2-AP for 6 h. IL-8 levels in cell culture supernatants were measured by ELISA. Data are expressed as mean ± SEM ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The levels of TSLP were measured using a mouse TSLP ELISA kit (catalog #MTLP00; R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s protocol.

Techniques: Western Blot, Phospho-proteomics, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Blockade of PAR2 Signaling by Punicalagin as a Therapeutic Strategy for Atopic Dermatitis

doi: 10.3390/ijms26188920

Figure Lengend Snippet: PCG normalizes epidermal hyperplasia, inflammatory markers, and neuronal hypersensitivity in DNFB-induced atopic dermatitis. ( A ) Representative H&E-stained skin sections showing epidermal thickness on day 28. Scale bar = 100 µm. ( B ) Quantitative analysis of epidermal thickness measured from H&E-stained sections ( n = 5–6 mice per group). ( C ) Serum TSLP levels measured by ELISA. ND, not detected ( n = 5–6 mice per group). ( D ) Calcium imaging of dorsal root ganglion (DRG) neurons in response to PAR2-AP (5 µM) stimulation. Traces show [Ca 2+ ] i changes over time, with KCl (100 mM) used as a positive control for neuronal viability ( n ≥ 20 per group). Data are presented as mean ± SEM. *** p < 0.001 compared to DNFB group.

Article Snippet: The levels of TSLP were measured using a mouse TSLP ELISA kit (catalog #MTLP00; R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s protocol.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Imaging, Positive Control

Journal: medRxiv

Article Title: Inhaled LTI-03 for Idiopathic Pulmonary Fibrosis: A Randomized Dose Escalation Study

doi: 10.1101/2025.10.28.25338981

Figure Lengend Snippet: A one-tailed, unpaired, parametric t-test was performed on the changes from baseline (pre-dose) to Day 14 in analyte values for placebo vs. LTI-03 5 mg/day, LTI-03 10 mg/day or pooled LTI-03. Differences from placebo are represented as point estimates with 90% confidence intervals. A p value of ≤ 0.1 was considered statistically significant. N: total number of samples; sample sizes are varied due to missing samples or results below the limit of quantification. DBB = deep bronchial brushings; SP-D = surfactant protein D; %P-AKT = percent phospho-AKT = COL1A1 = collagen type 1 alpha chain1; CXCL7 = chemokine (C-X-C motif) ligand 7; GAL-7 = galectin 7; IL-11 = interleukin 11; TSLP = thymic stromal lymphopoietin.

Article Snippet: Protein isolation and preparation of DBB samples for ELISAs were performed as previously described ( ) according to manufacturer’s instructions for COL1A1 (#ab210966, abcam), CXCL7 (#LS-F4967-1, LS Bio), GAL7 (#DY1339, R&D Systems), IL-11 (#DY218, R&D Systems), TSLP (#DY1398, R&D Systems).

Techniques: One-tailed Test