Extended Data Fig. 8: Overexpression of RPL12 promotes starvation-induced ribophagy in Drosophila.
a, Age-induced impairment of climbing ability was tested in WT, RpL12, RpL11, Nufip, and Atg5 RNAi flies. Data are shown as mean ± s.d. (5 groups, n = 10 for each group). b-e, Pupation (b), pupa emergence rate (c), as well as lengths of pupa (d) and adult flies (e) were quantified in wild-type (WT), RpL12, RpL11, Nufip, and Atg5 RNAi flies. Data are shown as mean ± s.d. (b, c, 5 groups, n = 20 each; d, e, n = 20). f, Expression levels of RpL12-3×FLAG, RpL12P3N-E21L(M)- 3×FLAG and RpL13-3×FLAG proteins were analyzed by western blot. Data are representative of two independent experiments. g, Climbing abilities of the indicated flies were quantified under starvation conditions. Data are shown as mean ± s.d. (5 groups, n = 10 each). h, Overexpression of wild-type RpL12 mitigates the loss of climbing ability induced by Aβ42. Elav-Gal4 was used to drive overexpression of RpL12, RpL12 P3N-E21L and RpL13 in the fly central nervous system (CNS). Data are shown as mean ± s.d. (4 groups, n = 20 each). i, WT flies and flies overexpressing RpL12-3×FLAG were subjected to starvation for 0 or 4 h. Ribophagic activity was assessed by measuring ribosomal protein levels before and after starvation. j, Degradation ratios from (i) were quantified and presented as mean ± s.d. k, Cell lysates were immunoprecipitated with anti-FLAG agarose beads and analyzed with the indicated antibodies. l, Ribosome profiling was performed in WT and RpL12-OE flies under nutrient-rich conditions. m, n, The fractions isolated from (l) were analyzed by western blot using the indicated antibodies. Tubulin served as a loading control in immunoblot. MHC-Gal4 was used to drive the expression of relevant genes in indirect flight muscle (f, g, k). Data in (i-k, m, n) are representative of three independent experiments, n = 15 for each replicate in f, i-k. P values were calculated by two-tailed Student’s t tests (a-e, g, h, j). ***P < 0.001, **P < 0.01, *P < 0.05, ns: no significance. Exact P values, source numerical data, and unprocessed blots are provided in Source data.
