recombinant mouse il 1β il 1f2 protein  (R&D Systems)

Journal: Glia

Article Title: Inflammatory Mediators Both Directly and Indirectly Promote Microglial Proliferation

doi: 10.1002/glia.70150

Figure Lengend Snippet: Astrocytes promote microglial proliferation basally and in response to IL1ɑ and IL1β stimulation. (A) Schematic depiction for the culture of astrocytes, subsequent media conditioning, and microglial treatment. (B) Bar plot depicting the percentage of proliferative microglia present in UCM and ACM. (C) Bar plot depicting the fold change in percentage of proliferative microglia in response to each screened ACM. B: N = 18 wells from 6 independent cultures; (C): N = 3–5 independent microglial cultures with treatments in triplicate. Bars represent mean ± SEM. (B): Two‐tailed, unpaired t ‐test; C: One‐way ANOVA, Dunnett's post hoc comparing fold changes to mean ACM control; ** p < 0.01, **** p < 0.0001. BDNF, Brain‐derived neurotrophic factor; CM, Conditioned media; CSF, Colony stimulating factor; CX3CL1, Fractalkine; IFN, Interferon; IGF, Insulin‐like growth factor; IL, Interleukin; LPA, Lysophosphatidic acid; NGFβ, Nerve growth factor β; NRG1, Neuregulin 1; NTS, Neurotensin; PTN, Pleiotrophin; S1P, Sphingosine‐1 phosphate; SCF, Stem cell factor; UCM, Unconditioned media; VEGF, Vascular endothelial growth factor; WNT3a, Wingless‐related int‐3a.

Article Snippet: After 5 days in vitro, we treated microglia with the following factors at 10 ng/mL: βNGF (Gibco 450–34), BDNF (Gibco 450–02), colony stimulating factor‐2 (CSF2; Gibco 315–03), CSF3 (Gibco AF‐250‐05), fractalkine (CX3CL1; Gibco 300–31), IFNɣ (ThermoFisher Scientific BMS326), IGF1 (Gibco 250–19), IL17A (Gibco 210–17), IL1ɑ (R&D Systems 400‐ML‐005), IL1β (R&D Systems 401‐ML‐005), interleukin‐3 (IL3; Gibco 213–13), IL33 (Gibco 210–33), IL4 (Gibco 214–14), IL5 (Gibco 215–15), LPA (Sigma Aldrich 857128P), NTS (R&D Systems 1909), NRG1 (Gibco 100–03), pleiotrophin (PTN; R&D Systems 6580‐PL), SCF (Gibco 250–03), sphingosine‐1‐phosphate (R&D Systems 1370), tumor necrosis factor ɑ (TNFɑ; Gibco 315‐01A), VEGF (R&D Systems 493‐MV‐005), and WNT3A (Gibco 315–20).

Techniques: Two Tailed Test, Control, Derivative Assay

Journal: Glia

Article Title: Inflammatory Mediators Both Directly and Indirectly Promote Microglial Proliferation

doi: 10.1002/glia.70150

Figure Lengend Snippet: Astrocyte‐secreted CSF2 promotes microglial proliferation in response to IL1ɑ and IL1β stimulation. (A) Plot depicting the percentage of proliferative microglia following 10 ng/mL treatment with each elevated cytokine identified by cytokine assay in microglia media lacking CSF1 and TGFβ2. (B) Plot depicting the concentration of CSF2 protein as determined by ELISA in 50× concentration ACM, IL1ɑ and IL1β ACM. (C) Representative images of 50× concentrated IL1ɑ ACM and IL1β ACM induced microglial proliferation (red = CD11b; white = KI67) and CSF2 neutralization. (D) Plot depicting the percentage of proliferative microglia in response to 50× concentrated UCM, ACM, IL1ɑ ACM with either IgG2A isotype control or ɑCSF2 treatment. (E) Plot depicting the percentage of proliferative microglia in response to 50× concentrated UCM, ACM, IL1β ACM with either IgG2A isotype control or ɑCSF2 treatment. Bars represent mean ± SEM. A: N = 12 wells from 3 independent cultures; (B): N = 4 batches of ACM; (D‐E): N = 9 wells from 3 independent cultures, the same AUCM and ACM controls were used for each of these experiments as they were prepared and tested alongside each respective IL1 ACM. (A, C, D): Colors represent independent microglial cultures: Large data points = mean percentage of proliferative microglia per culture, small data points = percentage of proliferative microglia per well. (A, B, D, E): One‐way ANOVA, Tukey's post hoc to compare differences between all conditions, * p < 0.05. Scale bars = 50 μm. ACM, Astrocyte conditioned media; CSF, Colony stimulating factor; CXCL, C‐X‐C motif chemokine ligand; CX3CL1, Fractalkine; IL, Interleukin; UCM, Unconditioned media.

Article Snippet: After 5 days in vitro, we treated microglia with the following factors at 10 ng/mL: βNGF (Gibco 450–34), BDNF (Gibco 450–02), colony stimulating factor‐2 (CSF2; Gibco 315–03), CSF3 (Gibco AF‐250‐05), fractalkine (CX3CL1; Gibco 300–31), IFNɣ (ThermoFisher Scientific BMS326), IGF1 (Gibco 250–19), IL17A (Gibco 210–17), IL1ɑ (R&D Systems 400‐ML‐005), IL1β (R&D Systems 401‐ML‐005), interleukin‐3 (IL3; Gibco 213–13), IL33 (Gibco 210–33), IL4 (Gibco 214–14), IL5 (Gibco 215–15), LPA (Sigma Aldrich 857128P), NTS (R&D Systems 1909), NRG1 (Gibco 100–03), pleiotrophin (PTN; R&D Systems 6580‐PL), SCF (Gibco 250–03), sphingosine‐1‐phosphate (R&D Systems 1370), tumor necrosis factor ɑ (TNFɑ; Gibco 315‐01A), VEGF (R&D Systems 493‐MV‐005), and WNT3A (Gibco 315–20).

Techniques: Cytokine Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Neutralization, Control

Journal: bioRxiv

Article Title: ER Stress-Induced β-Cell Apoptosis is Linked to Novel Select Lipid Signaling at the Transcriptional Level: Implications in T1D Developmen t

doi: 10.64898/2026.03.02.708596

Figure Lengend Snippet: MIN-6 β-cells were treated with cytokines, as in and in the absence or presence of P BA, BA Y 11, or A ZD for 16h. Cytosols were then prepared and processed for immunoblotting analyses for p65-NFκb, iPLA β, pSTAT1, pPERK, and CHOP. (Cytokine groups significantly different from corresponding DMSO groups, a p < 0.0001 and b p = 0.0003; c,d,*,e Cytokine + intervention groups significantly different from corresponding cytokine groups, p < 0.05, p < 0.01, p < 0.05 (1-tailed), and p < 0.005, respectively.

Article Snippet: Cells were treated with DMSO or cytokines (50 U/ml IL-1β + 150 U/ml IFNγ) (401-ML and 485-MI; R & D Systems, Minneapolis, MN) and processed for immunoblotting, TUNEL, ChIP, and lipidomics analyses.

Techniques: Western Blot

Journal: bioRxiv

Article Title: ER Stress-Induced β-Cell Apoptosis is Linked to Novel Select Lipid Signaling at the Transcriptional Level: Implications in T1D Developmen t

doi: 10.64898/2026.03.02.708596

Figure Lengend Snippet: MIN-6 β-cells were treated with D MSO or cytokines for 24h, as in , in the absence or presence of D MSO, P BA, S -BEL, F KGK18, R -BEL, C ay10502, scr ambled RNA, or si RNA targeting Pla2g6 . Cytosols were then prepared and processed for immunoblotting analyses for p65-NFκB, iPLA 2 β, pSTAT1, pPERK, and CHOP, and IκBα immunoblotting analyses. (Significance differences are presented as inserts.)

Article Snippet: Cells were treated with DMSO or cytokines (50 U/ml IL-1β + 150 U/ml IFNγ) (401-ML and 485-MI; R & D Systems, Minneapolis, MN) and processed for immunoblotting, TUNEL, ChIP, and lipidomics analyses.

Techniques: Western Blot

Journal: bioRxiv

Article Title: ER Stress-Induced β-Cell Apoptosis is Linked to Novel Select Lipid Signaling at the Transcriptional Level: Implications in T1D Developmen t

doi: 10.64898/2026.03.02.708596

Figure Lengend Snippet: A. MIN6 β-cells were pretreated with S -B EL (1 µM), FK GK18 (5 x 10 -8 M), R -B EL (1 µM), or Cay10502 (10 µM) for 1h or transfected with scr ambled or siRNA targeting Pla2g6 and then treated with D MSO or c ytokines (50 U/ml IL-1β + 150 U/ml IFNγ) for 24h and processed for TUNEL analyses. Data are means±SEMs of % apoptotic cells, relative to total cells. ( a Cytokine group significantly different from DMSO group, p<0.0001; b,c,d,e,f,g,h Cytokine group significantly different from cytokines + inhibitor or siRNA groups: p=0.0001, p=0.0002, p=0.0228, p=0.0007, p=0.0010, p=0.0004, p=0.0001, respectively.

Article Snippet: Cells were treated with DMSO or cytokines (50 U/ml IL-1β + 150 U/ml IFNγ) (401-ML and 485-MI; R & D Systems, Minneapolis, MN) and processed for immunoblotting, TUNEL, ChIP, and lipidomics analyses.

Techniques: Transfection, TUNEL Assay

Journal: bioRxiv

Article Title: ER Stress-Induced β-Cell Apoptosis is Linked to Novel Select Lipid Signaling at the Transcriptional Level: Implications in T1D Developmen t

doi: 10.64898/2026.03.02.708596

Figure Lengend Snippet: Human islets (500/condition) were treated with cytokines (100 U/ml IL-1β + 300 U/ml IFNγ) for 2-36h. In some replicates, at peak response times, cytokines + S -BEL groups were added. A. PGE 2 . B. PGF 2 α. C. 8-Iso-PGF 2 α. (Insets, *,† significantly different from DMSO group, p < 0.05 and p < 0.01, respectively, n=10-18. *CTK+ S -BEL group significantly different from CTK group, p < 0.05, n values indicated above the bars.)

Article Snippet: Cells were treated with DMSO or cytokines (50 U/ml IL-1β + 150 U/ml IFNγ) (401-ML and 485-MI; R & D Systems, Minneapolis, MN) and processed for immunoblotting, TUNEL, ChIP, and lipidomics analyses.

Techniques:

Journal: bioRxiv

Article Title: ER Stress-Induced β-Cell Apoptosis is Linked to Novel Select Lipid Signaling at the Transcriptional Level: Implications in T1D Developmen t

doi: 10.64898/2026.03.02.708596

Figure Lengend Snippet: MIN6 β-cells were treated with C ytokines ± inhibitors as in , or PGs, as in , and processed for iPLA 2 β immunoblotting analyses. Representative blots and corresponding densitometries for CTK ± S -BEL (A), CTK ± FKGK18, CTK ± R -BEL, CTK ± S -BEL, and cytokines and PGs (E) are presented. Cumulate densitometries are presented in and .

Article Snippet: Cells were treated with DMSO or cytokines (50 U/ml IL-1β + 150 U/ml IFNγ) (401-ML and 485-MI; R & D Systems, Minneapolis, MN) and processed for immunoblotting, TUNEL, ChIP, and lipidomics analyses.

Techniques: Western Blot

Journal: bioRxiv

Article Title: ER Stress-Induced β-Cell Apoptosis is Linked to Novel Select Lipid Signaling at the Transcriptional Level: Implications in T1D Developmen t

doi: 10.64898/2026.03.02.708596

Figure Lengend Snippet: Human islets (3500-5000/condition) from healthy donors were treated with cytokines (100 U/ml IL-1β + 300 U/ml IFNγ for 2h ( A, B ) or a PGs cocktail ( C ) as in for 4h. The islets were then processed for ChIP analyses using antibodies directed against p65-NFκB or iPLA 2 β and subsequent qPCR analyses. D . Human islets (250/condition) from healthy donors were treated with cytokines (100 U/ml IL-1β + 300 U/ml IFNγ or a PGs cocktail for 24h and processed for TUNEL analyses. Data are mean ± SEMs of %apoptotic cells, relative to total cells. ( a,b Significantly different from DMSO group, p < 0.0001 and p = 0.004, respectively.)

Article Snippet: Cells were treated with DMSO or cytokines (50 U/ml IL-1β + 150 U/ml IFNγ) (401-ML and 485-MI; R & D Systems, Minneapolis, MN) and processed for immunoblotting, TUNEL, ChIP, and lipidomics analyses.

Techniques: TUNEL Assay