cst 91882  (Cell Signaling Technology Inc)

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: Materials Today Bio

Article Title: Intravesical folate-conjugated hydroxyethyl starch micelles for pH-triggered co-delivery of epirubicin and TLR7 agonist toward synergistic chemoimmunotherapy of bladder cancer

doi: 10.1016/j.mtbio.2026.102835

Figure Lengend Snippet: (A) CLSM images of RAW 264.7 cells after treatment with LPS + IFN-γ and IL-4. Cell nuclei were stained with DAPI (blue), CD86 fluorescence displayed in green and CD206 fluorescence displayed in red. Scale bar: 10 μm; (B) Confocal imaging of CD86 and CD206 expression in M2-TAMs treated with free IMQ, FA-HES-EPI, and FA-HES-EPI/IMQ micelles. Cell nuclei were stained with DAPI (blue), CD86 fluorescence displayed in green and CD206 fluorescence displayed in red. Scale bar: 10 μm; (C) FCM analysis of expression of CD86 and CD206 after co-culturing M2-TAMs with different formulations; (D) Mean fluorescence intensity of CD86 and CD206 in M2-TAMs treated with different formulations. (n = 3), n.s.: p > 0.05, **p < 0.01and ****p < 0.0001 vs. the IL-4 group; ##p < 0.01and ####p < 0.0001 vs. the IL-4 group; (E) Expression levels of IL-6 and TNF-α in RAW264.7 cells treated with free IMQ and FA-HES-EPI/IMQ micelles. (n = 3), n.s.: p > 0.05, ***p < 0.001 and ****p < 0.0001; ####p < 0.0001; (F) Western blot analysis of TLR7/MyD88/NF-κB signaling in RAW 264.7 cells treated with free IMQ or FA-HES-EPI/IMQ micelles.

Article Snippet: Rabbit anti-CD86 polyclonal antibody, rabbit anti-CD206 polyclonal antibody, FITC goat anti-rabbit antibody, IgG/Alexa Fluor555 goat anti-rabbit antibody, APC anti-mouse CD206 antibody, PE anti-mouse CD86 antibody, purified anti-mouse CD16/32 antibody, intracellular Fixation/Permeabilization buffer kit, IL-6 ELISA kit and TNF-α ELISA kit were all purchased from Elabscience Biotechnology (Wuhan, China).

Techniques: Staining, Fluorescence, Imaging, Expressing, Western Blot