Perfection v33 scanner

The protein level was determined, according to the method of Tan et al. [31 (link)]. The antibodies used in this study included: PPARα (Absin, Shanghai, China), CPT1 (Proteintech, USA), FXR (Bioss, Beijing, China), HNF1α (Abcam, Cambridge, UK), GRP78 (Cell Signaling Technology, CST, Danvers, MA, USA), p-PERK (Bioss, Beijing, China), p-EIF2α (CST, Danvers, MA, USA), JNK (CST, Danvers, MA, USA), p-JNK (CST, Danvers, MA, USA), P38 (CST, Danvers, MA, USA), p-P38 (CST, Danvers, MA, USA), ERK(CST, Danvers, MA, USA), p-ERK(CST, Danvers, MA, USA), c-Jun (CST, Danvers, MA, USA), p-c-Jun (CST, Danvers, MA, USA), GAPDH (ZSGB-Bio, Beijing, China), FLAG (CST, Danvers, MA, USA), HA (CST, Danvers, MA, USA) and GFP (CST, Danvers, MA, USA). An electrochemiluminescence kit (Beyotime, Beijing, China) was used to visualize the immunoreactive protein, which was then scanned with an Epson Perfection V33 scanner.

For MCF-7, AU565, MDA-MB-231, and MDA-MB-468, 1 × 105 cells per well (5 × 10⁴ for A549, 4 × 10⁵ for BT-474, and 6 × 10⁴ for MCF-12A) were plated in 24-well plates and infected at 0.1–10 pfu/cell in 500 μL of appropriate medium. Then, 2 h postinfection, medium was replaced by 1 mL fresh medium and incubated up to 7 days. Medium was aspirated and cells fixed with 10% buffered formalin followed (VWR, Radnor, PA, USA) with 1% crystal violet (Sigma-Aldrich, St. Louis, MO, USA) staining. After staining was completed, the plates were scanned at high resolution using a perfection V33 scanner (Epson, Suwa, Japan). Images were converted to 8-bit on ImageJ version 1.53 t (http:imagej.nih.gov/ij, accessed on 3 January 2023), and the same area selection was applied to each well to measure the mean intensity. The maximal survivability was set for the average of each control untreated well, at each time point, and compared to other treated conditions to determine the percentage of remaining viability.

The rice florets of cv. YK17 and tms5 mutant plants were collected during the anthesis period. Mature pollen grains were stained using 1% I₂–KI solution and images were obtained using a DM1000 microscope (Leica, Germany). Rice florets were photographed using a Perfection V33 scanner (Epson, Japan).