Journal: Inflammation

Article Title: Red Blood Cell-Derived Exosomes Deliver Complement C5 to Exacerbate Neuroinflammation and Neuronal Injury after Intracerebral Hemorrhage

doi: 10.1007/s10753-026-02456-z

Figure Lengend Snippet: ICH-Exos increased necroptosis and M1 polarization in hemin-induced primary murine microglia. ( A ) Cell viability was determined using the CCK-8 assay. ( B ) Cell apoptosis was assessed via flow cytometry. ( C ) Western blot analysis was conducted to measure p-RIPK1, RIPK1, p-RIPK3, RIPK3, p-MLKL, and MLKL levels. ( D and E ) Flow cytometry was used to examine the proportions of CD86 + and CD163 + macrophages. ( F ) qRT-PCR analysis of iNOS, COX-2, CCL2, ARG1, CD206, and YM1 levels. ( G ) iNOS, COX-2, ARG1, and CD206 proteins levels. ( H ) ELISA was conducted to assess IL-1β, IL-6, and TNF-α levels. n = 3. * p < 0.05 vs. Control group; # p < 0.05 vs. Hemin group

Article Snippet: The mouse TNF-α (CSB-E04741m, CUSABIO, China), human TNF-α (KE00154, Proteintech), IL-1β (KE10003, KE00021, Proteintech), IL-6 (CSB-E04639m, CSB-E04638h, CUSABIO), and C5a (CSB-E08512h, CUSABIO) ELISA kits were operated according to the manufacturer’s instructions.

Techniques: CCK-8 Assay, Flow Cytometry, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Control

Journal: Inflammation

Article Title: Red Blood Cell-Derived Exosomes Deliver Complement C5 to Exacerbate Neuroinflammation and Neuronal Injury after Intracerebral Hemorrhage

doi: 10.1007/s10753-026-02456-z

Figure Lengend Snippet: ICH-Exos increased necroptosis of macrophages/microglia and M1 polarization after ICH. ( A ) Cell apoptosis was detected by TUNEL. ( B ) p-RIPK1, RIPK1, p-RIPK3, RIPK3, p-MLKL, and MLKL protein levels. ( C and D ) Immunofluorescence was used to detect IBA1 + iNOS + and IBA1 + ARG1 + . ( E ) iNOS, COX-2, CCL2, ARG1, CD206, and YM1 mRNA levels. ( F ) iNOS, COX-2, ARG1, and CD206 levels were analyzed by western blot. ( G ) ELISA was conducted to examine IL-1β, IL-6, and TNF-α. n = 5. * p < 0.05 vs. Sham group; # p < 0.05 vs. ICH group

Article Snippet: The mouse TNF-α (CSB-E04741m, CUSABIO, China), human TNF-α (KE00154, Proteintech), IL-1β (KE10003, KE00021, Proteintech), IL-6 (CSB-E04639m, CSB-E04638h, CUSABIO), and C5a (CSB-E08512h, CUSABIO) ELISA kits were operated according to the manufacturer’s instructions.

Techniques: TUNEL Assay, Immunofluorescence, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Inflammation

Article Title: Red Blood Cell-Derived Exosomes Deliver Complement C5 to Exacerbate Neuroinflammation and Neuronal Injury after Intracerebral Hemorrhage

doi: 10.1007/s10753-026-02456-z

Figure Lengend Snippet: ICH-Exos C5 promoted necroptosis and M1 polarization in hemin-induced microglia. ( A ) The heat map shows the differentially expressed proteins. ( B ) C5a level in brain tissue analyzed by IHC. * p < 0.05 vs. sham group. ( C ) C5a level in serum was detected by ELISA. * p < 0.05 vs. sham group. Under RIPA lysis conditions, the results of Western blot for C5a in ( D ) human RBC-Exos and ( E ) murine RBC-Exos after untreated EVs or after treatment with proteinase K (5 U/mL, 10 min) . # p < 0.05 vs. Sham-Exos/Normal-Exos group; @ p < 0.05 vs. ICH-Exos group. ( F ) C5a level in HMC3 cells was analyzed by Western blot and ELISA. ( G ) Cell viability was detected by CCK-8. ( H ) Cell apoptosis was detected by flow cytometry. ( I ) p-RIPK1, RIPK1, p-RIPK3, RIPK3, p-MLKL, and MLKL levels were detected by Western blot. ( G ) The proportions of CD86 + M1 and CD163 + M2 macrophages were detected by flow cytometry. ( K ) iNOS, COX-2, CCL2, ARG1, CD206, and YM1 levels analyzed by qRT-PCR. ( L ) iNOS, COX-2, ARG1 and CD206 levels analyzed by Western blot. (M) ELISA was used to detect IL-1β, IL-6, and TNF-α levels. n = 3. # p < 0.05 vs. Normal-Exos group; & p < 0.05 vs. ICH-Exos group

Article Snippet: The mouse TNF-α (CSB-E04741m, CUSABIO, China), human TNF-α (KE00154, Proteintech), IL-1β (KE10003, KE00021, Proteintech), IL-6 (CSB-E04639m, CSB-E04638h, CUSABIO), and C5a (CSB-E08512h, CUSABIO) ELISA kits were operated according to the manufacturer’s instructions.

Techniques: Paraffin-embedded Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Lysis, Western Blot, CCK-8 Assay, Flow Cytometry, Quantitative RT-PCR

Journal: Bioorganic & medicinal chemistry

Article Title: Inhibition of LPS-induced inflammatory response in RAW264.7 cells by natural Chlorogenic acid isomers involved with AKR1B1 inhibition.

doi: 10.1016/j.bmc.2024.117942

Figure Lengend Snippet: Fig. 6. DARTS-based determination of affinity between CGA isomers and AKR1B1. Graph representing the change in Anti-hydrolytic stability of intracellular AKR1B1 after treatment with (a) 3-CQA, (b) 4-CQA, and (c) 5-CQA for 15 min. The concentration of AKR1B1 was determined using an ELISA kit. Data are represented as mean ± SD, n = 3. Statistically significant differences between treated cells and control group were determined by the student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: TNF-α and IL-6 ELISA kit was purchased from Elabscience (Elabscience Biotechnology Co.,Ltd.

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

Journal: Bioorganic & medicinal chemistry

Article Title: Inhibition of LPS-induced inflammatory response in RAW264.7 cells by natural Chlorogenic acid isomers involved with AKR1B1 inhibition.

doi: 10.1016/j.bmc.2024.117942

Figure Lengend Snippet: Fig. 10. CETSA-based determination of target engagement between CGA isomers and AKR1B1. Graph representing the change in thermal stability of intracellular AKR1B1 after treatment with (a) 3-CQA, (b) 4-CQA, and (c) 5-CQA for 2 h. The concentration of AKR1B1 was determined using an ELISA kit. Data are represented as mean ± SD, n = 3. Statistically significant differences between treated cells and non-treated cells were determined by the student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: TNF-α and IL-6 ELISA kit was purchased from Elabscience (Elabscience Biotechnology Co.,Ltd.

Techniques: Drug discovery, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Cell Death & Disease

Article Title: Chromatin accessibility analysis identifies the transcription factor ETV5 as a suppressor of adipose tissue macrophage activation in obesity

doi: 10.1038/s41419-021-04308-0

Figure Lengend Snippet: A BMDMs were transfected with siRNA negative control (siRNA NC) or siRNA specific to Etv5 (siRNA Etv5 ) 48 h and Il6 expression was assessed by RT-qPCR 48 h later. Il6 expression levels of Raw264.7 cells after knockdown of Etv5 using shRNA B or overexpression of Etv5 C . Il6 expression levels of Raw264.7 cells after knockdown of Etv5 using shRNA D or overexpression of Etv5 E under MMe induction condition. F Il6 expression levels of BMDMs after treatment with palmitate, glucose, and insulin alone or with a combination of these compounds for 24 h. G–I Measuring IL-6 protein levels in the supernatant of Raw264.7 cell culture by ELISA. Cells were treated with palmitate (P) or PGI G , or underwent Etv5 knockdown using siRNA H , or shRNA I . Data in this result were all representative of three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, Student’s t test.

Article Snippet: Supernatants were collected after macrophage culture and IL-6 protein concentrations were measured using a mouse IL-6 ELISA kit (E-MSEL-M0001, Elabscience, China) according to the manufactories’ instructions.

Techniques: Transfection, Negative Control, Expressing, Quantitative RT-PCR, Knockdown, shRNA, Over Expression, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: ATF6 ameliorates renal warm ischemia-reperfusion injury through FHL2-mediated NF-κB signaling pathway

doi: 10.1016/j.isci.2026.115173

Figure Lengend Snippet: ATF6 plays a protective role in renal ischemia-reperfusion injury (A–B) The levels of serum Cr and BUN in mice. (C–D) The levels of serum IL-6 and TNF-α in mice. (E–F) The representative images and kidney injury assessment score of HE staining of the kidney. Ceapin-A7 (10 μM) is an ATF6 inhibitor, and AA147 (10 μM) is an ATF6 agonist. Data represent mean ± SDs, and statistical analysis was performed using one-way ANOVA. (∗ p < 0.05).

Article Snippet: IL-6 ELISA kit , Boster , EK0412.

Techniques: Staining

Journal: iScience

Article Title: ATF6 ameliorates renal warm ischemia-reperfusion injury through FHL2-mediated NF-κB signaling pathway

doi: 10.1016/j.isci.2026.115173

Figure Lengend Snippet: ATF6 plays a protective role in renal ischemia-reperfusion injury (A–B) The levels of serum Cr and BUN in mice. (C–D) The levels of serum IL-6 and TNF-α in mice. (E–F) The representative images and kidney injury assessment score of HE staining of the kidney. (G–H) RT-qPCR analysis of TNF-α and IL-6 expression in cultured HK-2 in response to H/R treatment. (I–J) RT-qPCR analysis of IL-6 and TNF-α expression in cultured HK-2 in response to H/R using siATF6. Data represent mean ± SDs, and statistical analysis was performed using one-way ANOVA. (∗ p < 0.05).

Article Snippet: IL-6 ELISA kit , Boster , EK0412.

Techniques: Staining, Quantitative RT-PCR, Expressing, Cell Culture