Journal: Scientific Reports

Article Title: Altered fibroblast-like synoviocyte epigenetics is responsible for deficient NUB1 expression in rheumatoid arthritis

doi: 10.1038/s41598-026-38420-y

Figure Lengend Snippet: Immunohistochemical staining of NUB1, NEDD8, IL-6, NF-kB in RA and OA synovial tissues. ( a ) Representative images (magnification, ×200) of immunohistochemistry for NUB1, NEDD8, and IL-6 in synovial tissues from patients with RA and OA. Expression of NEDD8 and IL-6 was higher in the intimal lining of RA synovium, and expression of NUB1 was lower compared with OA (see Fig. for quantification). Scale bar = 100 μm, ( b ) Representative images (magnification, ×400) of immunohistochemistry for NUB1 and p65 in synovial tissues from patients with RA and OA. In RA, regions with reduced NUB1 expression showed prominent nuclear localization of p65, whereas in OA, areas with higher NUB1 expression displayed weaker p65 nuclear staining. Negative control (NC) sections were stained with rabbit IgG under identical conditions. Scale bar = 50 μm.

Article Snippet: For immunohistochemistry, anti-NEDD8 (19E3) rabbit mAb (#2754), and anti-p65 rabbit mAb (#8242), were purchased from Cell Signaling Technology, and a rabbit polyclonal anti-NUB1 antibody (#14343-1-AP), and anti-rabbit polyclonal anti-IL-6 antibody (#21865-1-AP) were purchased from Proteintech, and normal Rabbit IgG control (#AB-105-C) was purchased from R༆D).

Techniques: Immunohistochemical staining, Staining, Immunohistochemistry, Expressing, Negative Control

Journal: Scientific Reports

Article Title: Altered fibroblast-like synoviocyte epigenetics is responsible for deficient NUB1 expression in rheumatoid arthritis

doi: 10.1038/s41598-026-38420-y

Figure Lengend Snippet: Immunohistochemical scoring of NUB1, NEDD8, and IL-6 in RA and OA synovial tissues. Synovial tissue sections from RA (n = 5) and OA (n = 5) patients were immunostained for NUB1, NEDD8, IL-6, and CCL5. For each patient, three randomly selected fields were evaluated in the lining and sublining zones. Staining was semiquantitatively scored as described in Methods. Graphs show the total scores as well as the average scores for the lining and sublining zones. For the total scores, NUB1 was higher in OA than in RA, whereas NEDD8 and IL-6 were significantly higher in RA; these differences were most prominent in the lining layer. Data are presented as mean ± SEM. Statistical analysis was performed using unpaired t test with Welch’s correction or Mann-Whitney U test (* p <0.05; ** p <0.01).

Article Snippet: For immunohistochemistry, anti-NEDD8 (19E3) rabbit mAb (#2754), and anti-p65 rabbit mAb (#8242), were purchased from Cell Signaling Technology, and a rabbit polyclonal anti-NUB1 antibody (#14343-1-AP), and anti-rabbit polyclonal anti-IL-6 antibody (#21865-1-AP) were purchased from Proteintech, and normal Rabbit IgG control (#AB-105-C) was purchased from R༆D).

Techniques: Immunohistochemical staining, Staining, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Altered fibroblast-like synoviocyte epigenetics is responsible for deficient NUB1 expression in rheumatoid arthritis

doi: 10.1038/s41598-026-38420-y

Figure Lengend Snippet: Regulation of NUB1 expression by MAPKs . RA and OA FLS (n = 5 each) were stimulated with IL-1β (2 ng/mL) for 6h in the presence or absence of MAPK inhibitors: SP600125 (JNK inhibitor), SB203580 (p38 MAPK inhibitor), and U0126 (MEK1/2–ERK inhibitor). NUB1 mRNA expression is shown as fold change relative to IL-1β stimulation alone. ( a ) Quantitative analysis demonstrated that none of the MAPK inhibitors significantly reduced IL-1β–induced NUB1 mRNA expression in either RA or OA FLS (IL-1β vs. IL-1β + SB203580, adjusted p > 0.9999; IL-1β vs. IL-1β + SP600125, adjusted p > 0.9999; IL-1β vs. IL-1β + U0126, adjusted p = 0.8285). ( b ) IL-6 mRNA expression was quantified by RT-qPCR and normalized to GAPDH. IL-6 expression was significantly reduced by each MAPK inhibitor compared with IL-1β stimulation alone (**** p < 0.0001), confirming effective inhibition of MAPK-dependent signaling. Data are presented as mean ± SEM. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test.

Article Snippet: For immunohistochemistry, anti-NEDD8 (19E3) rabbit mAb (#2754), and anti-p65 rabbit mAb (#8242), were purchased from Cell Signaling Technology, and a rabbit polyclonal anti-NUB1 antibody (#14343-1-AP), and anti-rabbit polyclonal anti-IL-6 antibody (#21865-1-AP) were purchased from Proteintech, and normal Rabbit IgG control (#AB-105-C) was purchased from R༆D).

Techniques: Expressing, Quantitative RT-PCR, Inhibition

Journal: Skeletal Muscle

Article Title: TRPV1 manipulating polarization of M1/M2 macrophages to promote skeletal muscle regeneration

doi: 10.1186/s13395-026-00417-6

Figure Lengend Snippet: TRPV1 activation modulate protein expression of markers and cytokines associated with M1 and M2 in vivo, respectively. A Representative Western blot of CD86(M1); Arg1(M2). B-E The protein expression or mRNA level of CD86 and the protein expression of IL-6(M1) ( n = 3 animals per experimental group; mean ± SD; Two-way ANOVA). F , G The protein expression of Arg1 ( n = 3 animals per experimental group; mean ± SD; Two-way ANOVA). H , I The mRNA level or protein expression of CD206(M2) ( n = 3 animals per experimental group; mean ± SD; Two-way ANOVA). Statistical significance was set at P < 0.05. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: For cytological immunofluorescence, cells were fixed in 4% paraformaldehyde, permeabilized with 0.5% Triton X-100 for 15 min. After blocking with 5% BSA, the slides were incubated with the primary antibody for TRPV1, CD86, CD206, mouse anti-MYH3 (1:200, Santa Cruz Biotechnology, sc-53091), rabbit anti-IL-6 polyclonal antibody (1:200, Proteintech, #21865-1-AP), and rabbit anti-Arginase-1 polyclonal antibody (1:200, Proteintech, #16001-1-AP).

Techniques: Activation Assay, Expressing, In Vivo, Western Blot

Journal: Skeletal Muscle

Article Title: TRPV1 manipulating polarization of M1/M2 macrophages to promote skeletal muscle regeneration

doi: 10.1186/s13395-026-00417-6

Figure Lengend Snippet: TRPV1 inhibits M1 polarization and promotes M2 polarization in vitro. A , C Representative western blot of CD86(M1); Arg1(M2). B , D The expression of CD86 and Arg-1 in response to LPS/IFN-γ and IL-4/IL-10 stimulation at the protein level, respectively ( n = 3 independent replicates in cells; mean ± SD; One-way ANOVA). E Representative Western blot of IL-6 and CD86(M1) after treatment with CAP and CPZ in vitro. F-I The protein expression of IL-6 and CD86 in different groups ( n = 3 independent replicates in cells; mean ± SD; One-way ANOVA). J Representative western blot of CD206 and Arg1 (M2) after treatment with CAP and CPZ in vitro. K-N The protein expression of CD206 and Arg1 in each group ( n = 3 independent replicates in cells; mean ± SD; One-way ANOVA). Statistical significance was set at P < 0.05. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: For cytological immunofluorescence, cells were fixed in 4% paraformaldehyde, permeabilized with 0.5% Triton X-100 for 15 min. After blocking with 5% BSA, the slides were incubated with the primary antibody for TRPV1, CD86, CD206, mouse anti-MYH3 (1:200, Santa Cruz Biotechnology, sc-53091), rabbit anti-IL-6 polyclonal antibody (1:200, Proteintech, #21865-1-AP), and rabbit anti-Arginase-1 polyclonal antibody (1:200, Proteintech, #16001-1-AP).

Techniques: In Vitro, Western Blot, Expressing