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: p-RIPK1 , 28252-1-AP , Rabbit , 1:1000 , 70–80 KDa , Proteintech , China.

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: p-RIPK1 , 28252-1-AP , Rabbit , 1:1000 , 70–80 KDa , Proteintech , China.

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: p-RIPK1 , 28252-1-AP , Rabbit , 1:1000 , 70–80 KDa , Proteintech , China.

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