Ribosomal Protein S6/RPS6 Antibody

Detection of RPS6 by Western Blot

Tor1 N-terminal HEAT repeats were required to accelerate growth and activate TORC1 in response to preferred nitrogen sources and to phosphate.A. Cells of indicated genotypes were grown in YNB without ammonium sulfate ((NH4)2SO4) supplemented with 10 mM Glutamine (Gln), 38 mM (NH4)2SO4, 10 mM Proline (Pro) or 10 mM Tryptophan (Trp) as sole nitrogen source, without or with 300 ng/ml doxycycline (300 Doxy). (TOR1/TOR1, JKC1713; tor1/TOR1, JKC1347; tetO-TOR1-Del381, JKC1441; tetO-TOR1, JKC1549) B. Western blot of cells pre-grown in YPD medium with 2 μg/ml doxycycline for 4 h (Time 0), then incubated in YNB without doxycycline, without (NH4)2SO4, with 10 mM Proline (Pro) or Glutamine (Gln) as sole nitrogen source for 45 min; protein extracts probed with antibody to phosphorylated Rps6 (P-S6), total Rps6 (S6) or phosphorylated eukaryotic translation initiation factor 2A (P-eIF2a), and tubulin (Tub) as loading control. Dens: signal intensity ratio of P-S6 or P-eIF2a to Tub. Strains as in panel B and TOR1/TOR1, JKC1361. Same samples were run on separate gels to detect either P-S6 or P-eIF2a. C. Cells pre-grown as in A were inoculated into YNB without phosphate supplemented with varying concentration of KH2PO4 (Pi), without or with 300 ng/ml doxycycline (300 Doxy). Error bars show SD of 3 technical replicates. Strains as in panel A and tor1/TOR1, JKC1347. Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.ppat.1010089), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of RPS6 by Western Blot

The HOPS complex is required for the activation of TOR pathway.a Rapamycin sensitivity examined by spot assay. C. albicans strains were serial 5-fold diluted, spotted on YPD agar containing rapamycin and cultured at 30 °C for 48 h. b The phosphorylation of RPS6 protein (P-S6) examined by western blotting. The representative images shown are from 3 biological repeats. c The hyphal morphology of C. albicans strains on solid medium. Three biological replicates were carried out. One of the representative images are shown. Width (W) and depth (D) (mm) were quantified by Image J. n = 5 colonies. Scale bars = 1 mm. d Relative expression of GTR1 and TOR1 in GTR1OE-vam6 delta / delta mutants compared to SC5314 examined by quantitative real-time RT-PCR. n = 3 samples. e Rapamycin sensitivity examined by spot assay. f The ratio of germination in C. albicans cultured in hyphal-inducing liquid media at 37 °C for 0.5 h. The ratio of germinated strains to all the strains observed in the microscopic field (approximately 50–100 fungal cells in each field under the 40× objective) was considered as the ratio of germination. n = 3 fields. g The hyphal length quantified by Image J. n = 50 hyphae. h The hyphal morphology of C. albicans strains on solid medium. Three biological replicates were carried out. One of the representative images are shown. Width (W) and depth (D) (mm) were quantified by Image J. Scale bars = 1 mm. n = 3 colonies. Data were presented as mean ± SD (c, d, f, h). Box plots indicate median (middle line), 25th, 75th percentile (box) and minimum and maximum value as well as outliers (single points) (g). Two-tailed unpaired t-test (c, d, f, g, h). Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41467-024-48525-5), licensed under a CC-BY license. Not internally tested by R&D Systems.