RT-PCR Confirmation of Mutant Strains

To confirm the mutant strain Δstk and the complementary strain CΔstk, we also performed RT-PCR (Tan et al., 2015 (link)). Briefly, RNA was isolated using SV Total RNA Isolation System (Promega, USA) according to the manufacturer's instructions. Then, cDNA was synthesized using HiScript Q Select RT SuperMix (Vazyme, China) according to the manufacturer's instructions.
To confirm whether the upstream and downstream genes of stk in the mutant were affected or not, the genes were amplified using the primers SSU05_0427-F/SSU05_0427-R (for upstream gene) and SSU05_0429-F/SSU05_0429-R (for downstream gene; Table S1) using the SC-19 cDNA as template.