Fig. 5: CTRP9 gene knockout reduces CMA activity by downregulating LAMP2A expression during both myoblast differentiation and in mature C2C12 myotubes.
From: CTRP9 as a myokine mitigates sarcopenia via the LAMP-2A/NLRP3 pathway
A Western blot analysis of LC3A and LC3B expression in siNC and siCTRP9 groups (n = 3). B Western blot analysis of LAMP2A, LAMP-1, and HSC70 expression in siNC and siCTRP9 groups; GAPDH serves as the loading control (n = 3). C The mRNA expression levels of LAMP2A in siNC and siCTRP9 groups. (n = 3). D Western blot analysis of NLRP3, pro-caspase-1, IL-1β, LAMP2A, and LAMP-1 expression in gastrocnemius muscle from 3-month-old and 23-month-old WT and CTRP9 KO mice; GAPDH serves as the loading control (n = 5). E Immunofluorescence staining of siNC and siCTRP9 C2C12 myotubes for NLRP3 (green), LAMP2A (red), and DAPI-stained nuclei (blue) (n = 3). Scale bar = 20 μm. F Western blot analysis of NLRP3, LAMP-1, LAMP2A and IL-1β expression in siNC and siCTRP9 C2C12 myoblasts; GAPDH serves as the loading control (n = 3). G Immunofluorescence staining of siNC and siCTRP9 C2C12 myoblasts for NLRP3 (green), LAMP2A (red), and DAPI-stained nuclei (blue) (n = 3). Scale bar = 20 μm. Data are expressed as mean ± SEM.
