Journal: Materials Today Bio

Article Title: Apoptotic extracellular vesicles derived from MSCs exposed to hypoxic and inflammatory environments slow intervertebral disc degeneration by enhancing cell activity and regulating immunity microenvironment

doi: 10.1016/j.mtbio.2026.103013

Figure Lengend Snippet: A) Immunofluorescence staining of macrophage polarization markers CD86 and CD206 in macrophages treated with different ApoEVs in vitro (scale bar = 25 μm) and B) corresponding quantitative analysis. C-E) qRT-PCR analysis was performed to evaluate the gene expression of M1 macrophage surface markers (iNOS, TNF-a and IL-1β) in macrophages cultured with different ApoEVs. F-G, I) qRT-PCR analysis was performed to evaluate the gene expression of M2 macrophage surface markers (Arg-1, IL-10 and CD163) in macrophages cultured with different ApoEVs. H) Immunofluorescence staining of M1 macrophage surface markers (CD86) and M2 macrophage surface markers (CD206) in IVD tissues in vivo (scale bar = 1 mm). J) Quantitative analysis of immunofluorescence staining for CD206 and CD86 in IVD tissues in vivo. ns, no significance ( P > 0.05); * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Subsequently, cells were permeabilized with 0.1% Triton X-100 (Thermo Fisher Scientific, America) for 15 min, followed by blocking with immunostaining blocking solution (Beyotime, China) for 1 h. Primary antibodies targeting P53 (Beyotime, China), MMP13 (Bioss, China), COL II (Bioss, China), ACAN (Bioss, China), CD86 (proteintech, China), CD206 (proteintech, China), BCL-2 (Beyotime, China) and γ-H2AX (Beyotime, China) were applied and incubated overnight at 4 °C.

Techniques: Immunofluorescence, Staining, In Vitro, Quantitative RT-PCR, Gene Expression, Cell Culture, In Vivo

Journal: Materials Today Bio

Article Title: Apoptotic extracellular vesicles derived from MSCs exposed to hypoxic and inflammatory environments slow intervertebral disc degeneration by enhancing cell activity and regulating immunity microenvironment

doi: 10.1016/j.mtbio.2026.103013

Figure Lengend Snippet: A) Immunofluorescence staining of STAT6 (signal transducer and activator of transcription 6) in macrophages (scale bar = 50 μm) and B) corresponding quantitative analysis. C) Immunofluorescence staining and D) corresponding quantitative analysis of STAT6 in macrophages treated with or without specific STAT6 antagonist (AS1517499). E) β-gal staining of NPCs in different groups (scale bar = 100 μm). F) Semi-quantitative analysis of SA-β-gal positive cells in different groups. G) Immunofluorescence staining of ROS in NPCs (scale bar = 50 μm). H) Cell viability of NPCs during senescence induction was detected by the CCK-8 assay. I, J) qRT-PCR analysis of iNOS and TNF-α in macrophages cultured with different ApoEVs. K, L) qRT-PCR analysis of Arg-1 and IL-10 in macrophages. M) Immunofluorescence staining of CD86 and CD206 in macrophages (scale bar = 25 μm) and P) corresponding quantitative analysis. N) Safranin O staining of NPCs in different media for 3 days (scale bar = 100 μm). O) Immunofluorescence staining of COL II and TNF-α in each group (scale bar = 100 μm) and Q, R) corresponding quantitative analysis. The AS1517499 group means the I-ApoEVs treatment group with the addition of a specific STAT6 antagonist (AS1517499). ns, no significance ( P > 0.05); * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Subsequently, cells were permeabilized with 0.1% Triton X-100 (Thermo Fisher Scientific, America) for 15 min, followed by blocking with immunostaining blocking solution (Beyotime, China) for 1 h. Primary antibodies targeting P53 (Beyotime, China), MMP13 (Bioss, China), COL II (Bioss, China), ACAN (Bioss, China), CD86 (proteintech, China), CD206 (proteintech, China), BCL-2 (Beyotime, China) and γ-H2AX (Beyotime, China) were applied and incubated overnight at 4 °C.

Techniques: Immunofluorescence, Staining, CCK-8 Assay, Quantitative RT-PCR, Cell Culture

Journal: Materials Today Bio

Article Title: Apoptotic extracellular vesicles derived from MSCs exposed to hypoxic and inflammatory environments slow intervertebral disc degeneration by enhancing cell activity and regulating immunity microenvironment

doi: 10.1016/j.mtbio.2026.103013

Figure Lengend Snippet: A) Representative T2-weighted MRI images of rat coccygeal vertebrae at 4 weeks after surgery and different treatment. B) Pfirrmann grade analysis was used to quantitatively evaluate the disc degeneration degree. C) The changes of DHI in each group were calculated by X-ray images. D) X-ray images of the intervertebral disc of rat coccygeal at 4 weeks after treatment. E) Immunofluorescence staining of CD86 and CD206 in IVD tissues in vivo (scale bar = 500 μm) and F) corresponding quantitative analysis. G) TNF-α and COL II co-immunofluorescence staining of nucleus pulposus tissue in the intervertebral disc in vivo (scale bar = 500 μm) and H) corresponding quantitative analysis. The AS1517499 group means the I-ApoEVs treatment group with the addition of a specific STAT6 antagonist (AS1517499). ns, no significance ( P > 0.05); * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Subsequently, cells were permeabilized with 0.1% Triton X-100 (Thermo Fisher Scientific, America) for 15 min, followed by blocking with immunostaining blocking solution (Beyotime, China) for 1 h. Primary antibodies targeting P53 (Beyotime, China), MMP13 (Bioss, China), COL II (Bioss, China), ACAN (Bioss, China), CD86 (proteintech, China), CD206 (proteintech, China), BCL-2 (Beyotime, China) and γ-H2AX (Beyotime, China) were applied and incubated overnight at 4 °C.

Techniques: Immunofluorescence, Staining, In Vivo

Journal: Kidney International Reports

Article Title: Pauci-Immune Endocapillary Proliferative Glomerulonephritis With Glomerular M2 Macrophage Infiltration

doi: 10.1016/j.ekir.2026.103791

Figure Lengend Snippet: Multiplex immunofluorescence staining results (Case 3). (a) Macrophage subtypes and their distribution patterns. (b) CD86+ macrophages are sparsely distributed within the renal interstitium. (c–e) CD163+ and CD206+ macrophages are abundantly present in the interstitium, whereas CD163+ macrophages show less infiltration within the glomeruli. The scale bar in the figure represents 20 μm. DAPI, 4',6-diamidino-2-phenylindole.

Article Snippet: Formalin-fixed paraffin-embedded renal tissue sections (3.5 μm) were subjected to multiplex IF staining using the following primary antibodies: CD68 (ab955, pan-macrophage marker, Abcam), CD163 (ab182422, M2 macrophage marker, Abcam), CD86 (91882S, M1 macrophage marker, Cell Signaling Technology, Danvers, MA), CD206 (24595S, M2 macrophage marker, Cell Signaling Technology), CD56 (99746S, Cell Signaling Technology), CD3 (ab11089, Abcam), and CD8 (ab199016, Abcam).

Techniques: Multiplex Assay, Immunofluorescence, Staining