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KIF1B inhibition reduces IL‐13‐induced inflammatory responses in BEAS‐2B cells. (A) BEAS‐2B cells were transfected with si‐KIF1B or si‐NC and treated with IL‐13 (10 ng/mL) for 24 h, followed by Western blot analysis of NLRP3, cleaved caspase‐1 and cleaved GSDMD protein expression. Conditioned medium was collected to measure cytokine levels: <t>(B)</t> <t>TNF‐α,</t> (C) IL‐1β, (D) IL‐18 and (E) IL‐10 by ELISA. β‐actin was used as the loading control for normalisation in Western blot. Data are presented as mean ± SD ( n = 3; *** p < 0.001 vs. control, p < 0.01, p < 0.001 vs. IL‐13 group).

Murine Balf, supplied by R&D Systems, used in various techniques. Bioz Stars score: 96/100, based on 1435 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "KIF1B Regulates NLRP3 ‐Mediated Pyroptosis in Asthma Progression"

Article Title: KIF1B Regulates NLRP3 ‐Mediated Pyroptosis in Asthma Progression

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.70975

Figure Legend Snippet: KIF1B inhibition reduces IL‐13‐induced inflammatory responses in BEAS‐2B cells. (A) BEAS‐2B cells were transfected with si‐KIF1B or si‐NC and treated with IL‐13 (10 ng/mL) for 24 h, followed by Western blot analysis of NLRP3, cleaved caspase‐1 and cleaved GSDMD protein expression. Conditioned medium was collected to measure cytokine levels: (B) TNF‐α, (C) IL‐1β, (D) IL‐18 and (E) IL‐10 by ELISA. β‐actin was used as the loading control for normalisation in Western blot. Data are presented as mean ± SD ( n = 3; *** p < 0.001 vs. control, p < 0.01, p < 0.001 vs. IL‐13 group).

Techniques Used: Inhibition, Transfection, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Control

KIF1B regulates IL‐13‐induced inflammation in BEAS‐2B cells via NLRP3. BEAS‐2B cells were transfected with si‐NC, si‐KIF1B + empty vector or si‐KIF1B + NLRP3‐ORF plasmid, with or without IL‐13 treatment. (A) NLRP3 gene expression was measured by qRT‐PCR following NLRP3‐ORF transfection to confirm successful overexpression. (B) Following transfection with si‐KIF1B with or without NLRP3‐ORF and IL‐13 treatment, intracellular ROS, MDA and GSH levels were measured. (C) Protein expression of NLRP3, cleaved caspase‐1 and cleaved GSDMD was assessed by Western blot. (D) Cytokine levels (TNF‐α, IL‐1β, IL‐18 and IL‐10) in conditioned medium were measured by ELISA. GAPDH gene was used as an internal reference for qPCR analysis, and β‐actin was used as the loading control for normalisation in Western blot. Data are presented as mean ± SD ( n = 3; *** p < 0.001 vs. control, p < 0.01, p < 0.001 vs. IL‐13 group, & p < 0.05, & p < 0.01 vs. IL‐13 + si‐KIF1B group).

Figure Legend Snippet: KIF1B regulates IL‐13‐induced inflammation in BEAS‐2B cells via NLRP3. BEAS‐2B cells were transfected with si‐NC, si‐KIF1B + empty vector or si‐KIF1B + NLRP3‐ORF plasmid, with or without IL‐13 treatment. (A) NLRP3 gene expression was measured by qRT‐PCR following NLRP3‐ORF transfection to confirm successful overexpression. (B) Following transfection with si‐KIF1B with or without NLRP3‐ORF and IL‐13 treatment, intracellular ROS, MDA and GSH levels were measured. (C) Protein expression of NLRP3, cleaved caspase‐1 and cleaved GSDMD was assessed by Western blot. (D) Cytokine levels (TNF‐α, IL‐1β, IL‐18 and IL‐10) in conditioned medium were measured by ELISA. GAPDH gene was used as an internal reference for qPCR analysis, and β‐actin was used as the loading control for normalisation in Western blot. Data are presented as mean ± SD ( n = 3; *** p < 0.001 vs. control, p < 0.01, p < 0.001 vs. IL‐13 group, & p < 0.05, & p < 0.01 vs. IL‐13 + si‐KIF1B group).

Techniques Used: Transfection, Plasmid Preparation, Gene Expression, Quantitative RT-PCR, Over Expression, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Control

KIF1B knockdown reduces OVA‐induced inflammation and pyroptosis in mouse pulmonary tissues. C57BL/6 mice were infected with lentiviral particles carrying sh‐KIF1B or sh‐NC, and subsequently treated with OVA for 14 days to induce asthmatic responses. Lung tissues were collected at sacrifice. (A) Wright‐Giemsa staining was used to quantify total cells, macrophages, eosinophils, lymphocytes and neutrophils in BALF. (B) IgE, TNF‐α, IL‐1β, IL‐18 and IL‐10 levels in BALF were measured by ELISA. (C) Protein levels of NLRP3, cleaved caspase‐1 and cleaved GSDMD in lung homogenates were assessed by Western blot. β‐actin was used as the loading control for normalisation in Western blot. Data are presented as mean ± SD ( n = 8; *** p < 0.001 vs. control, p < 0.01, p < 0.001 vs. OVA group).

Figure Legend Snippet: KIF1B knockdown reduces OVA‐induced inflammation and pyroptosis in mouse pulmonary tissues. C57BL/6 mice were infected with lentiviral particles carrying sh‐KIF1B or sh‐NC, and subsequently treated with OVA for 14 days to induce asthmatic responses. Lung tissues were collected at sacrifice. (A) Wright‐Giemsa staining was used to quantify total cells, macrophages, eosinophils, lymphocytes and neutrophils in BALF. (B) IgE, TNF‐α, IL‐1β, IL‐18 and IL‐10 levels in BALF were measured by ELISA. (C) Protein levels of NLRP3, cleaved caspase‐1 and cleaved GSDMD in lung homogenates were assessed by Western blot. β‐actin was used as the loading control for normalisation in Western blot. Data are presented as mean ± SD ( n = 8; *** p < 0.001 vs. control, p < 0.01, p < 0.001 vs. OVA group).

Techniques Used: Knockdown, Infection, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Control

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Journal: Journal of Cellular and Molecular Medicine

Article Title: KIF1B Regulates NLRP3 ‐Mediated Pyroptosis in Asthma Progression

doi: 10.1111/jcmm.70975

Figure Lengend Snippet: KIF1B inhibition reduces IL‐13‐induced inflammatory responses in BEAS‐2B cells. (A) BEAS‐2B cells were transfected with si‐KIF1B or si‐NC and treated with IL‐13 (10 ng/mL) for 24 h, followed by Western blot analysis of NLRP3, cleaved caspase‐1 and cleaved GSDMD protein expression. Conditioned medium was collected to measure cytokine levels: (B) TNF‐α, (C) IL‐1β, (D) IL‐18 and (E) IL‐10 by ELISA. β‐actin was used as the loading control for normalisation in Western blot. Data are presented as mean ± SD ( n = 3; *** p < 0.001 vs. control, p < 0.01, p < 0.001 vs. IL‐13 group).

Article Snippet: For murine samples (BALF), TNF‐α (Cat# MTA00B, R&D Systems), IL‐1β (Cat# MLB00C, R&D Systems), IL‐18 (Cat# DY7625‐05, R&D Systems) and IL‐10 (Cat# M1000B, R&D Systems) were assessed.

Techniques: Inhibition, Transfection, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: KIF1B Regulates NLRP3 ‐Mediated Pyroptosis in Asthma Progression

doi: 10.1111/jcmm.70975

Figure Lengend Snippet: KIF1B regulates IL‐13‐induced inflammation in BEAS‐2B cells via NLRP3. BEAS‐2B cells were transfected with si‐NC, si‐KIF1B + empty vector or si‐KIF1B + NLRP3‐ORF plasmid, with or without IL‐13 treatment. (A) NLRP3 gene expression was measured by qRT‐PCR following NLRP3‐ORF transfection to confirm successful overexpression. (B) Following transfection with si‐KIF1B with or without NLRP3‐ORF and IL‐13 treatment, intracellular ROS, MDA and GSH levels were measured. (C) Protein expression of NLRP3, cleaved caspase‐1 and cleaved GSDMD was assessed by Western blot. (D) Cytokine levels (TNF‐α, IL‐1β, IL‐18 and IL‐10) in conditioned medium were measured by ELISA. GAPDH gene was used as an internal reference for qPCR analysis, and β‐actin was used as the loading control for normalisation in Western blot. Data are presented as mean ± SD ( n = 3; *** p < 0.001 vs. control, p < 0.01, p < 0.001 vs. IL‐13 group, & p < 0.05, & p < 0.01 vs. IL‐13 + si‐KIF1B group).

Techniques: Transfection, Plasmid Preparation, Gene Expression, Quantitative RT-PCR, Over Expression, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: KIF1B Regulates NLRP3 ‐Mediated Pyroptosis in Asthma Progression

doi: 10.1111/jcmm.70975

Figure Lengend Snippet: KIF1B knockdown reduces OVA‐induced inflammation and pyroptosis in mouse pulmonary tissues. C57BL/6 mice were infected with lentiviral particles carrying sh‐KIF1B or sh‐NC, and subsequently treated with OVA for 14 days to induce asthmatic responses. Lung tissues were collected at sacrifice. (A) Wright‐Giemsa staining was used to quantify total cells, macrophages, eosinophils, lymphocytes and neutrophils in BALF. (B) IgE, TNF‐α, IL‐1β, IL‐18 and IL‐10 levels in BALF were measured by ELISA. (C) Protein levels of NLRP3, cleaved caspase‐1 and cleaved GSDMD in lung homogenates were assessed by Western blot. β‐actin was used as the loading control for normalisation in Western blot. Data are presented as mean ± SD ( n = 8; *** p < 0.001 vs. control, p < 0.01, p < 0.001 vs. OVA group).

Techniques: Knockdown, Infection, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Demographics and clinical characteristics of IPF patients.

Journal: Frontiers in Medicine

Article Title: Increased lipocalin-2 expression in pulmonary inflammation and fibrosis

doi: 10.3389/fmed.2023.1195501

Figure Lengend Snippet: Demographics and clinical characteristics of IPF patients.

Article Snippet: LCN2 levels were estimated in human and murine BALF using a commercially available ELISA kit (EA100541, OriGene Technologies Inc.), according to the manufacturer's instructions.

Techniques:

LCN2 BALF levels of IPF patients negatively correlate with their respiratory functions. (A–C) Spearman's correlation plots of LCN2 levels, as measured using a commercially available ELISA, with the ratio of forced expiratory volume (FEV)/forced vital capacity (FVC) (A) , with a transfer capacity of the lung for the uptake of carbon monoxide (TLCO) (B) and with carbon monoxide transfer coefficient (KCO) (C) . Statistical significance was assessed with Spearman's r = −0.47, −0.42 as indicated; * denotes p < 0.05.

Journal: Frontiers in Medicine

Article Title: Increased lipocalin-2 expression in pulmonary inflammation and fibrosis

doi: 10.3389/fmed.2023.1195501

Figure Lengend Snippet: LCN2 BALF levels of IPF patients negatively correlate with their respiratory functions. (A–C) Spearman's correlation plots of LCN2 levels, as measured using a commercially available ELISA, with the ratio of forced expiratory volume (FEV)/forced vital capacity (FVC) (A) , with a transfer capacity of the lung for the uptake of carbon monoxide (TLCO) (B) and with carbon monoxide transfer coefficient (KCO) (C) . Statistical significance was assessed with Spearman's r = −0.47, −0.42 as indicated; * denotes p < 0.05.

Article Snippet: LCN2 levels were estimated in human and murine BALF using a commercially available ELISA kit (EA100541, OriGene Technologies Inc.), according to the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay

Increased Lcn2 expression in mouse lungs during the development of BLM-induced pulmonary inflammation and fibrosis. (A) Total protein concentration in BALFs, as determined using the Bradford assay. (B) Inflammatory cell numbers in BALFs, as counted with a hemacytometer. (C) Soluble collagen levels in the BALFs as detected with the Direct Red assay. Statistical significance was assessed with one-way ANOVA; */** denote p < 0.05/0.01 respectively. (D) Representative images from H&E-stained lung sections of murine lungs at 3, 7, and 14 d post-BLM administration (×10). (E–J) Respiratory functions were measured with FlexiVent, 14 days post-BLM; mean respiratory system compliance (Crs); mean respiratory system elastance (Ers); mean tissue elastance (H); mean static lung compliance (Cst); mean total lung capacity (A). Cumulative results from three independent experiments; statistical significance was assessed using the Mann–Whitney test; */** denote p < 0.05/0.01, respectively. (K) Lcn2 mRNA expression was interrogated using Q-RT-PCR; Values were normalized over the expression of the housekeeping gene B2m and presented as fold change over control. (L, M) Lcn2 concentration in BALF (L) and serum (M) of mice at 3, 7, and 14 d post-BLM administration. Lcn2 levels were measured using a commercially available ELISA kit; Statistical significance was assessed with one-way ANOVA, */**/*** denote p < 0.05/0.01/0.001 respectively. (N) Bar plot showing the percentage of immune cell populations in the murine lung post-BLM; the employed gating strategy is described in . (O) Representative Western blot of Lcn2 expression (red) in fibrotic lungs, 14 d post-BLM. (P) Densitometry analysis of Lcn2 expression, normalized to the expression of Actin (green); cumulative result from two independent experiments; statistical significance was assessed with unpaired t -test; ** denotes p < 0.01. (Q) Representative images of two independent experiments, from immunohistochemistry for Lcn2 in control (SAL) and fibrotic (BLM) murine lung tissue (×10).

Journal: Frontiers in Medicine

Article Title: Increased lipocalin-2 expression in pulmonary inflammation and fibrosis

doi: 10.3389/fmed.2023.1195501

Figure Lengend Snippet: Increased Lcn2 expression in mouse lungs during the development of BLM-induced pulmonary inflammation and fibrosis. (A) Total protein concentration in BALFs, as determined using the Bradford assay. (B) Inflammatory cell numbers in BALFs, as counted with a hemacytometer. (C) Soluble collagen levels in the BALFs as detected with the Direct Red assay. Statistical significance was assessed with one-way ANOVA; */** denote p < 0.05/0.01 respectively. (D) Representative images from H&E-stained lung sections of murine lungs at 3, 7, and 14 d post-BLM administration (×10). (E–J) Respiratory functions were measured with FlexiVent, 14 days post-BLM; mean respiratory system compliance (Crs); mean respiratory system elastance (Ers); mean tissue elastance (H); mean static lung compliance (Cst); mean total lung capacity (A). Cumulative results from three independent experiments; statistical significance was assessed using the Mann–Whitney test; */** denote p < 0.05/0.01, respectively. (K) Lcn2 mRNA expression was interrogated using Q-RT-PCR; Values were normalized over the expression of the housekeeping gene B2m and presented as fold change over control. (L, M) Lcn2 concentration in BALF (L) and serum (M) of mice at 3, 7, and 14 d post-BLM administration. Lcn2 levels were measured using a commercially available ELISA kit; Statistical significance was assessed with one-way ANOVA, */**/*** denote p < 0.05/0.01/0.001 respectively. (N) Bar plot showing the percentage of immune cell populations in the murine lung post-BLM; the employed gating strategy is described in . (O) Representative Western blot of Lcn2 expression (red) in fibrotic lungs, 14 d post-BLM. (P) Densitometry analysis of Lcn2 expression, normalized to the expression of Actin (green); cumulative result from two independent experiments; statistical significance was assessed with unpaired t -test; ** denotes p < 0.01. (Q) Representative images of two independent experiments, from immunohistochemistry for Lcn2 in control (SAL) and fibrotic (BLM) murine lung tissue (×10).

Article Snippet: LCN2 levels were estimated in human and murine BALF using a commercially available ELISA kit (EA100541, OriGene Technologies Inc.), according to the manufacturer's instructions.

Techniques: Expressing, Protein Concentration, Bradford Assay, Staining, MANN-WHITNEY, Reverse Transcription Polymerase Chain Reaction, Control, Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Immunohistochemistry

Lcn2 expression is upregulated during LPS-induced Acute Lung Injury (ALI). (A) Total protein concentration in BALF, as determined using the Bradford assay. (B) Inflammatory cell numbers in BALF, from saline and LPS-treated mice, as counted with a hemacytometer. (C) Lcn2 mRNA expression was interrogated with Q-RT-PCR; Values were normalized over the expression of the housekeeping gene B2m and presented as fold change over control; representative results from three independent experiments. (D) Western blot of Lcn2 expression (red) in lungs from mice with LPS-induced ALI, followed by densitometry analysis of Lcn2 expression, normalized to the expression of Actin (green). (E) Representative images from immunohistochemistry for Lcn2 in lungs from control (SAL) and LPS-treated mice (×10). (F, G) Lcn2 levels in BALF (F) and serum (G) of mice were estimated using ELISA; statistical significance was assessed using the Mann–Whitney test; */**** denote p < 0.05/0.0001.

Journal: Frontiers in Medicine

Article Title: Increased lipocalin-2 expression in pulmonary inflammation and fibrosis

doi: 10.3389/fmed.2023.1195501

Figure Lengend Snippet: Lcn2 expression is upregulated during LPS-induced Acute Lung Injury (ALI). (A) Total protein concentration in BALF, as determined using the Bradford assay. (B) Inflammatory cell numbers in BALF, from saline and LPS-treated mice, as counted with a hemacytometer. (C) Lcn2 mRNA expression was interrogated with Q-RT-PCR; Values were normalized over the expression of the housekeeping gene B2m and presented as fold change over control; representative results from three independent experiments. (D) Western blot of Lcn2 expression (red) in lungs from mice with LPS-induced ALI, followed by densitometry analysis of Lcn2 expression, normalized to the expression of Actin (green). (E) Representative images from immunohistochemistry for Lcn2 in lungs from control (SAL) and LPS-treated mice (×10). (F, G) Lcn2 levels in BALF (F) and serum (G) of mice were estimated using ELISA; statistical significance was assessed using the Mann–Whitney test; */**** denote p < 0.05/0.0001.

Article Snippet: LCN2 levels were estimated in human and murine BALF using a commercially available ELISA kit (EA100541, OriGene Technologies Inc.), according to the manufacturer's instructions.

Techniques: Expressing, Protein Concentration, Bradford Assay, Saline, Reverse Transcription Polymerase Chain Reaction, Control, Western Blot, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

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