MARK4 Antibody - #AF0693

Fig. 4. TNTs inhibit MARK4 expression to decrease tubulin glutamylation and NLRP3 inflammasome activation. (A) Western blot analysis of MARK4 expression in macrophages cultured on pTi and TNTs surfaces under LPS and Nigericin stimulation. (B) Immunofluorescence images showing MARK4 expression in macrophages cultured on pTi and TNTs surfaces. 3D intensity plots illustrate fluorescence distribution. Scale bars, 10 μm. (C) Western blot analysis of MARK4 and inflammasome-related proteins including cell lysates and supernatants in BMDMs with MARK4 inhibitor-1 treatment. (D–E) ELISA and LDH release assays showing the effects of MARK4 inhibitor-1 on IL-1β secretion and pyroptosis. (F) Western blot analysis of MARK4 and inflammasome-related proteins including cell lysates and supernatants in BMDMs overexpressing MARK4 via adenoviral transduction. (G–H) ELISA and LDH release assays showing the effects of MARK4 overexpression on IL-1β secretion and pyroptosis. (I) Western blot of MARK4 protein levels in BMDMs after siCCP5 and siTTLL4 knockdown. (J) Western blot of microtubule glutamylation levels following MARK4 overexpression and MARK4 inhibitor-1 knockdown. (K) Immunofluorescence staining of glutamylated tubulin (green), total tubulin (red), and nuclei (blue) in macrophages cultured following MARK4 overexpression and MARK4 inhibitor-1 knockdown. Scale bars, 10 μm. (L–M) ELISA quantification of IL-1β secretion in BMDMs following combined manipulations of MARK4 expression and tubulin glutamylation-related factors. (L) Knockdown of MARK4 via shRNA reversed the increase in IL-1β secretion induced by siCCP5. (M) Overexpression of MARK4 via adenovirus rescued the reduction in IL-1β secretion caused by siTTLL4. (N) Western blot analysis of NLRP3 inflammasome components and glutamylated tubulin levels in macrophages treated with MARK4 inhibitor-1 or siCCP5. The data are presented as the mean ± SD of three biological replicates. ∗p < 0.05, ∗∗p < 0.01, and ns (not significant). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 4. TNTs inhibit MARK4 expression to decrease tubulin glutamylation and NLRP3 inflammasome activation. (A) Western blot analysis of MARK4 expression in macrophages cultured on pTi and TNTs surfaces under LPS and Nigericin stimulation. (B) Immunofluorescence images showing MARK4 expression in macrophages cultured on pTi and TNTs surfaces. 3D intensity plots illustrate fluorescence distribution. Scale bars, 10 μm. (C) Western blot analysis of MARK4 and inflammasome-related proteins including cell lysates and supernatants in BMDMs with MARK4 inhibitor-1 treatment. (D–E) ELISA and LDH release assays showing the effects of MARK4 inhibitor-1 on IL-1β secretion and pyroptosis. (F) Western blot analysis of MARK4 and inflammasome-related proteins including cell lysates and supernatants in BMDMs overexpressing MARK4 via adenoviral transduction. (G–H) ELISA and LDH release assays showing the effects of MARK4 overexpression on IL-1β secretion and pyroptosis. (I) Western blot of MARK4 protein levels in BMDMs after siCCP5 and siTTLL4 knockdown. (J) Western blot of microtubule glutamylation levels following MARK4 overexpression and MARK4 inhibitor-1 knockdown. (K) Immunofluorescence staining of glutamylated tubulin (green), total tubulin (red), and nuclei (blue) in macrophages cultured following MARK4 overexpression and MARK4 inhibitor-1 knockdown. Scale bars, 10 μm. (L–M) ELISA quantification of IL-1β secretion in BMDMs following combined manipulations of MARK4 expression and tubulin glutamylation-related factors. (L) Knockdown of MARK4 via shRNA reversed the increase in IL-1β secretion induced by siCCP5. (M) Overexpression of MARK4 via adenovirus rescued the reduction in IL-1β secretion caused by siTTLL4. (N) Western blot analysis of NLRP3 inflammasome components and glutamylated tubulin levels in macrophages treated with MARK4 inhibitor-1 or siCCP5. The data are presented as the mean ± SD of three biological replicates. ∗p < 0.05, ∗∗p < 0.01, and ns (not significant). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Figure 1. Increased MARK4 and the NLRP3 inflammasome activation in inflamed gingival tissues (a) Gene transcription in the healthy (n = 10) and inflamed (n = 10) gingiva was measured by real time qPCR. (b and c) Samples from healthy (n = 6) and inflamed gingiva (n = 6) were stained by the immunolohistological chemistry. The scale bar represents 50 µm. (d) Protein levels in the gingival tissue from chronic periodontitis (CP, n = 3) or normal control (NC, n = 3) were evaluated by Western blot. *p < 0.05; **p < 0.01; ***p < 0.001.

Figure 1. Increased MARK4 and the NLRP3 inflammasome activation in inflamed gingival tissues (a) Gene transcription in the healthy (n = 10) and inflamed (n = 10) gingiva was measured by real time qPCR. (b and c) Samples from healthy (n = 6) and inflamed gingiva (n = 6) were stained by the immunolohistological chemistry. The scale bar represents 50 µm. (d) Protein levels in the gingival tissue from chronic periodontitis (CP, n = 3) or normal control (NC, n = 3) were evaluated by Western blot. *p < 0.05; **p < 0.01; ***p < 0.001.

Fig. 7 Theabrownin reduces MARK4 expression in HFD-fed obese mice. (a) Immunofluorescence of MARK4 in hippocampal CA1, CA3 regions, and cortex (Scale = 20 μm). (b)Immunofluorescence statistics of MARK4 in hippocampal CA1, CA3 regions, and cortex (n = 3). (c) Representative Western blot of MARK4. (d) Histogram representing the quantitative analysis of MARK4(n = 3). mean ± SEM. *p < 0.05, **p < 0.01.