human duo set elisa kits  (R&D Systems)

Journal: Redox Biology

Article Title: C-terminal interleukin 1 alpha (IL-1α) overexpression drives EMT and a vulnerability to ferroptosis in HNSCC

doi: 10.1016/j.redox.2026.104172

Figure Lengend Snippet: Characterization of IL-1α overexpression constructs in Cal27 HNSCC cells. Gene expression (A-C), protein release (D) and protein expression (E-G) of the three IL1A constructs – Full- Length (FL), N-terminal (NT) and C-terminal (CT) in the Cal27 IL-1α-overexpressing cells were analyzed by RT-qPCR, ELISA and Western blot respectively compared to control. GAPDH was used as an endogenous control for PCR analysis (A-C); α-Tubulin was used as the loading control for whole cell lysate and cytoplasmic fraction western blots (E-F); and Lamin B1 was used as the loading control for the nuclear fraction Western blot (G). Bars represent mean ± SEM from n = 3 independent experiments. IL-1α band intensities were normalized to the respective loading controls. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001.

Article Snippet: Cell culture media from IL-1α overexpressed Cal27 cells were collected for analyzing the levels of IL-1α, IL-1β, IL-6, IL-8 or IL-1RA using Human Duo Set ELISA kits (R&D Systems, Minneapolis, MN) according to the manufacturer's protocols.

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

Journal: Redox Biology

Article Title: C-terminal interleukin 1 alpha (IL-1α) overexpression drives EMT and a vulnerability to ferroptosis in HNSCC

doi: 10.1016/j.redox.2026.104172

Figure Lengend Snippet: IL-1α overexpression alters expression of IL-1 pathway genes. Gene expression of IL1B (A), IL6 (B), IL8 (C), IL1RN (D), IL1RAP (E) and IL1R1 (F) in the three IL1A constructs – Full-Length (FL), N-terminal (NT) and C-terminal (CT) in Cal27 IL-1αoverexpressing cells were analyzed by RT-PCR using GAPDH as an endogenous control. Protein secretion of IL-1β (G), IL-6 (H), IL-8 (I), and IL1RA (J) in cell culture supernatants was quantified by ELISA with protein concentrations normalized to cell numbers. Whole cell lysates were analyzed for IL-1R1 expression by Western blot using GAPDH as a loading control (K). Bars represent mean ± SEM from n = 3 independent experiments.∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001.

Article Snippet: Cell culture media from IL-1α overexpressed Cal27 cells were collected for analyzing the levels of IL-1α, IL-1β, IL-6, IL-8 or IL-1RA using Human Duo Set ELISA kits (R&D Systems, Minneapolis, MN) according to the manufacturer's protocols.

Techniques: Over Expression, Expressing, Gene Expression, Construct, Reverse Transcription Polymerase Chain Reaction, Control, Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Molecular Medicine Reports

Article Title: Veronicastrum sibiricum (L.) Pennell extract alleviates inflammation-induced muscle atrophy through NLRP3 inflammasome regulation and mitochondrial function restoration

doi: 10.3892/mmr.2026.13857

Figure Lengend Snippet: VSE reduces the secretion of proinflammatory cytokines. C2C12 myotubes were pretreated with VSE at 0 (LPS only), 1.25, 2.5 and 5 µg/ml for 3 h and then co-treated with 500 ng/ml LPS for 24 h. Controls received no treatments. (A) mRNA expression levels of the proinflammatory cytokines TNF-α and IL-1β were analyzed using reverse transcription-quantitative PCR. (B) Protein expression levels of the proinflammatory cytokines TNF-α and IL-1β in the supernatant of C2C12 myotube cultures were analyzed using ELISA. All data are presented as the mean ± SD of triplicate results and statistical analysis was carried out using one-way ANOVA with Tukey's post hoc test. #### P<0.0001 vs. control; ***P<0.001, ****P<0.0001 vs. LPS treatment. A450, absorbance at 450 nm; LPS, lipopolysaccharide; VSE, Veronicastrum sibiricum seed extract.

Article Snippet: Supernatants were acquired, and secretion of the proinflammatory cytokines TNF-α (cat. no. MTA00B; R&D systems, Inc.) and IL-1β was analyzed using ELISA kits (cat. no. DY401; R&D Systems, Inc.) according to the manufacturer's guidelines.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Control

Journal: bioRxiv

Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

doi: 10.64898/2026.04.21.719931

Figure Lengend Snippet: ( a ) Cell viability was assessed with the MTT assay and data shown are means ± SD (n = 3). ( b ) TNFα, IL-6 and IL-8 expression in response to B[a]P exposure. ( c ) Representative Western blot images showing protein expression. All data were obtained from three independent experiments. Statistical p values were determined by a one-way ANOVA, followed by a Tukey post-hoc test comparing the B[a]P treatments to the DMSO control; * p < 0.05; ¥, p < 0.01; NS, no significance.

Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

Techniques: MTT Assay, Expressing, Western Blot, Control

Journal: bioRxiv

Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

doi: 10.64898/2026.04.21.719931

Figure Lengend Snippet: Cells were treated with various concentrations of PM2.5 for 24 hr. ( a ) Cell viability was assessed with MTT assay. ( b ) IL-6 and IL-8 from the basolateral sides. ( c ) Representative Western blot images showing CFTR, β2AR, phospho-AKT (Ser473), AKT, phospho-NF-κB p65 (Ser536), NF-κB p65, and TRPC6 expression. All data are presented as the mean ± SD (n = 4). Statistical p values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing the B[a]P or PM 2.5 treatments to the DMSO control; *, p < 0.05; ¥, p < 0.01; NS, no significance.

Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

Techniques: MTT Assay, Western Blot, Expressing, Control

Journal: bioRxiv

Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

doi: 10.64898/2026.04.21.719931

Figure Lengend Snippet: ( a ) IL-6 and IL-8 from the basolateral sides of the ALI were determined using the DuoSet® ELISA kits. Data are presented as the mean ± SD (n = 4). Statistical p -values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing the all treatments to the DMSO control; ¥, p < 0.05; *, p < 0.01. ( b ) Representative Western blot images showing CFTR, phospho-AKT (Ser473), AKT, phospho-NF-κB p65 (Ser536), and NF-κB p65 expression in treated polarized 16HBE14o-cells in ( a ). All data were obtained from three independent experiments.

Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

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

Journal: bioRxiv

Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

doi: 10.64898/2026.04.21.719931

Figure Lengend Snippet: ALI cells were treated with DMSO, 100 ng/mL PM 2.5, 10 µM LY 294002 ± PM 2.5, or 10 µM LY 303511 ± PM 2.5 for 24 hrs. ( a ) IL-6 and IL-8 from the basolateral sides were determined using the DuoSet® ELISA kits. Data are presented as the mean ± SD (n=3). Statistical p values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing all treatments to the DMSO control, or PM 2.5 to PM 2.5 ± LY 294002 or LY 303511; *, p < 0.01. ( b ) Representative Western blot images showing CFTR and β-actin expression. β-actin was used to ensure equal loading of protein. All data were obtained from three independent experiments, and each experiment was analyzed in duplication.

Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

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

Journal: bioRxiv

Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

doi: 10.64898/2026.04.21.719931

Figure Lengend Snippet: ( a ) The lung parenchyma from two ferrets were sliced into several sections as shown. Arrows indicate the tracheobronchial tubes. All tissue sections were incubating with DMSO (control), 10 µM B[a]P, or 100 ng/mL PM 2.5 in Pneumacult-EX Plus Basal media for 24 hrs. ( b ) IL 6 secretion was determined using the ferret ELISA kit. Data represent the mean of 6 independent sections. Statistical p values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing the B[a]P or PM 2.5 treatments to the DMSO control; ¥, p < 0.05 (n = 6). ( c ) Representative western blotting images show reduction of CFTR, and β2AR and elevation of TRPC6 expression in explants under these treatments.

Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

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