Figure 7

Lysosomal protein content and CMA substrate steady-state levels after prolonged starvation. Immunoblotting and quantitative RT-PCR of lysates from N2a cells either non-transfected (non) or stably transfected with scramble shRNA (scmbl) or shRNA targeting LAMP-2 mRNA. Cells were cultured in EBSS media for 24 hours to induce CMA. (a) Immunoblots and respective densitometric quantification showing protein levels of CMA substrates MEF2D and GAPDH (b) Immunoblots and respective densitometric quantification of LAMP-2A and (c) quantitative RT-PCR of LAMP-2A and LAMP-2B. (d) Immunoblots and respective densitometric quantification showing protein levels of LAMP-1. (e) Quantitative RT-PCR of LAMP-1. (f) Immunoblots and respective densitometric quantification showing protein levels of LIMP-2. (g) Quantitative RT-PCR of LIMP-2. (Samples were cultured in EBSS for 24 hours to induce CMA; actin was used to control loading; *p < 0.05, **p < 0.01, ***p < 0.001).