RNA-seq of H. dujardinii larval stage, rep. 3
H. dujardinii samples were collected at the end of June 2025 (6630.145 N, 3307.765 E) at a water temperature of 10C and a depth of up to 6 m. For differential gene expression analysis, the following were used: (1) free-swimming or in-tissue larvae; (2) adult sponge tissues; and (3) cell aggregates formed in vitro for 24 h from mechanically dissociated adult sponge cells. Each group was represented by at least three biological replicates. The quality of total RNA was evaluated using the Bioanalyzer 2100 (Agilent, Santa Clara, CA, USA.  500 ng of RIN 7 of total RNA was used for library construction using NEBNext Poly(A) mRNA Magnetic Isolation Module and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA, USA) according to the manufacturers instruction. The quality of libraries was verified using the Bioanalyzer 2100 (Agilent, Santa Clara, CA, USA) and the yield was validated by qPCR. Libraries were then sequenced on NovaSeq6000 (Illumina, San Diego, CA, USA) with pair-end 100 bp reading.