Additional file 1 of Granulin as an important immune molecule involved in lamprey tissue repair and regeneration by promoting cell proliferation and migration
Creators: name: Sun, Ruixiang
name: Wang, Dong
name: Song, Yuxuan
name: Li, Qingwei
name: Su, Peng
name: Pang, Yue
Publisher: name: figshare
Year: 2022
Subjects: subject: Genetics
subject: FOS: Biological sciences

description: Additional file 1: Fig S1. Amino acid and nucleotide sequence alignment of Pm-PGRN-S1 and Pm-PGRN-S2. A: Amino acid sequence alignment. B: Nucleotide sequence alignment. Fig S2. Phylogenetic tree analysis of PGRN domains in jawless and jawed mammals. Hs-PGRN is represented by the red background. Lr-PGRNs are represented by the blue background. Pm-PGRNs are represented by wathet-blue background. GRN A module: pink semiarc; GRN B module: atrovirens semiarc; GRN C module: blue semiarc; GRN D module: purple semiarc; GRN E module: light-red semiarc; GRN F module: wathet-blue semiarc; GRN G module: bottle-green semiarc. Fig S3. Collinear analysis of lamprey pgrns1-s3. The right side of the lamprey pgrn-s2 is the positive direction. Identical genes are represented by the same color. A semicircle–rectangle represents a gene, and identical genes are represented by the same color. The black line represents chromosome (Chr). Fig S4. Expression and purification of rL-PGRN-S1 protein. A: rL-PGRN-S1 protein expression results. M: protein marker; 1: uninduced expression of E. coli; 2–7: induced expression of E. coli. B: rL-PGRN-S1 protein purification results. M: protein marker; 1: uninduced expression of E. coli; 2: induced expression of E. coli; 3: supernatant; 4–5: inclusion body; 6: filtered; 7: binding elution; 8: elution with 400 mmoL/L imidazole; 9: 0.2 μg/μL BSA. Fig S5. Titer detection of Lr-PGRN-S1 polyclonal antibody using ELISA. Serially diluted (1:20,000–1:640,000) polyclonal antibodies were tested against Lr-PGRN-S1 recombinant protein by ELISA. Pre-immune IgG was used as negative control (n = 3). Error bars indicate standard error of the mean (s.e.m.). Fig S6. Expression and knockdown efficiency of Lr-pgrn genes during embryonic development. A: Expression of Lr-pgrn genes in gastrula and cephalic stages detected by qPCR. B: Knockdown efficiency of Lr-pgrn-s1 detected by qPCR. The probability of statistical differences between experimental groups was determined by Student’s t-test. Mean ± SD is shown (n = 3 per group). ns: nonsignificant, *P &lt; 0.05, **P &lt; 0.001, ***P &lt; 0.0001. Fig S7. Molecular changes in the MAPK, Notch, Wnt, and TGF-β signaling pathways in KEGG enrichment analysis of transcriptomic data. Green represents downregulated genes; red represents downregulated genes. Fig S8. A: GO annotation map of DEG. Red represents biological processes; Green represents cellular components; Blue represents molecular functions. B: KEGG classification map of differential gene. Cellular processes: yellow; environmental information processing: purple; genetic information processing: pink; human diseases: red; metabolism: green; organismal systems: blue. Table S1. Primers used for PCR in this study. Table S2. Primers used for siRNA and vector linking in this study.