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. 2019 Sep 8;16(7):1172–1185. doi: 10.1080/15548627.2019.1659573

Figure 1.

Figure 1.

Spt4 negatively regulates ATG8/Atg8 and ATG41/Atg41 expression in growing conditions. (A-D) Analysis of Atg8 and Atg41 protein levels. Protein extracts were generated from (A) wild-type (TVY1, pep4∆) and spt4∆ (WXY100, pep4∆) cells, or (C) wild-type (ZYY108) and spt4∆ (WXY114) cells expressing Atg41-PA, after growth in YPD to mid-log phase (growing conditions) and shifted to SD-N medium for 2 h (nitrogen starvation). Western blots were probed with anti-Atg8, anti-PA and anti-Pgk1 (loading control) antisera or antibodies. (B and D) Quantitative analysis of Atg8 and Atg41 protein levels, respectively. The protein level of either Atg8 or Atg41-PA after 2 h of starvation was set as 1 and other values were normalized; the error bar represents the SEM of at least 3 independent experiments. N.S., not significant, ***, p < 0.005. (E) Wild-type (WLY176) and spt4∆ (WXY105) cells were cultured until mid-log phase and collected in growing conditions. mRNA levels were quantified by RT-qPCR. The mRNA level of different ATG genes and YAT1 (negative control) in spt4∆ cells were normalized to that of the corresponding genes in wild-type cells, which was set as 1. Error bars represent the SEM of at least 3 independent experiments; two-tailed t test was used for statistical significance. p values are reported for the comparison between WT and spt4∆ strains in growing conditions. **, p < 0.01; ***, p < 0.005.