Journal: The Journal of Headache and Pain

Article Title: S100A11 regulates microglial inflammatory response in neuropathic pain via H3K27ac-TFEB-mitochondrial autophagy axis

doi: 10.1186/s10194-026-02328-9

Figure Lengend Snippet: S100A11 knockdown inhibits NLRP3-mediated pyroptosis in primary microglia. A Western blot analysis of NLRP3, GSDMD-N, ASC, and Caspase-1 protein levels of primary microglia; B-E Relative levels normalized to β-actin were quantified (for three independent experiments). F Western blot analysis of Bcl-2, Bax protein levels of primary microglia; G-H Relative levels normalized to β-actin were quantified (for three independent experiments). Immunofluorescence staining of NLRP3 ( I ) and GSDMD ( J ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels K , L , for three independent experiments; Immunofluorescence staining of Caspase1 ( M ) and ASC ( N ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels O , P , for three independent experiments; Q Staining of primary microglia with Yo-pro and PI fluorescent probes; nuclei were labeled with Hoechst. R Quantification plot of the mean fluorescence intensity of Yo-pro, for three independent experiments. Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001 . Statistical significance was determined by two-way ANOVA with appropriate post hoc tests

Article Snippet: Membranes were blocked with 5% skim milk for 60 min, rinsed with TBST, and incubated with primary antibodies at 4 °C overnight (β-actin: Proteintech, 66009-1-Ig, 1:10000; S100A11: Abcam, ab169530, 1:1000; NLRP3: AdipoGen, AG-20B-0014, 1:1000; GSDMD-N: Abcam, ab215203, 1:1000; ASC: Wanleibio, WL02462, 1:500; Caspase1: CST, 89332, 1:1000; Bax: Proteintech, 50599-2-Ig, 1:2000; Bcl-2: abclone, A19693, 1:2000; COXIV: Proteintech, 11242-1-AP, 1:1000; LC3: Proteintech, 14600-1-AP, 1:2000; P62: Proteintech, 18420-1-AP, 1:2000; Parkin: Proteintech, 14060-1-AP, 1:1000; Pink1: Proteintech, 23274-1-AP, 1:2000; H3: Proteintech, 17168-1-AP, 1:2000; TFEB: Proteintech, 13372-1-AP, 1:1000; TFEC: Proteintech, 13547-1-AP, 1:1000; TFE3: Proteintech, 14480-1-AP, 1:1000; H3K9ac: CST, 9649, 1:1000; H3K18ac: CST, 13998, 1:1000; H3K27ac: CST, 8173, 1:1000; H4K12ac; CST, 13944, 1:1000; H4K16ac: CST, 13534, 1:1000).

Techniques: Knockdown, Western Blot, Immunofluorescence, Staining, Fluorescence, Labeling

Journal: The Journal of Headache and Pain

Article Title: S100A11 regulates microglial inflammatory response in neuropathic pain via H3K27ac-TFEB-mitochondrial autophagy axis

doi: 10.1186/s10194-026-02328-9

Figure Lengend Snippet: S100A11 knockdown ameliorates neuronal dysfunction in the spinal cord of CCI model. A Schematic diagram of microglia collection and co-incubation with neurons. B-D Western blot analysis of Bcl-2, Bax protein levels of neurons; Relative levels normalized to β-actin were quantified (for three independent experiments). E Immunofluorescence staining of MAP-2 in primary neurons. F Staining of primary neurons with Calcein AM probe; G-I Western blot analysis of PSD-95 and SYP protein levels in L4-L6 spinal cord tissues of mice; Relative levels normalized to actin were quantified ( n = 3). L , M Representative immunofluorescence images of PSD-95, SYP expression in L4-L6 spinal cord segments of mice, and relative fluorescence intensity was quantified in panels J and K ( n = 3). Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001 . Statistical significance was determined by two-way ANOVA with appropriate post hoc tests

Article Snippet: Membranes were blocked with 5% skim milk for 60 min, rinsed with TBST, and incubated with primary antibodies at 4 °C overnight (β-actin: Proteintech, 66009-1-Ig, 1:10000; S100A11: Abcam, ab169530, 1:1000; NLRP3: AdipoGen, AG-20B-0014, 1:1000; GSDMD-N: Abcam, ab215203, 1:1000; ASC: Wanleibio, WL02462, 1:500; Caspase1: CST, 89332, 1:1000; Bax: Proteintech, 50599-2-Ig, 1:2000; Bcl-2: abclone, A19693, 1:2000; COXIV: Proteintech, 11242-1-AP, 1:1000; LC3: Proteintech, 14600-1-AP, 1:2000; P62: Proteintech, 18420-1-AP, 1:2000; Parkin: Proteintech, 14060-1-AP, 1:1000; Pink1: Proteintech, 23274-1-AP, 1:2000; H3: Proteintech, 17168-1-AP, 1:2000; TFEB: Proteintech, 13372-1-AP, 1:1000; TFEC: Proteintech, 13547-1-AP, 1:1000; TFE3: Proteintech, 14480-1-AP, 1:1000; H3K9ac: CST, 9649, 1:1000; H3K18ac: CST, 13998, 1:1000; H3K27ac: CST, 8173, 1:1000; H4K12ac; CST, 13944, 1:1000; H4K16ac: CST, 13534, 1:1000).

Techniques: Knockdown, Incubation, Western Blot, Immunofluorescence, Staining, Expressing, Fluorescence