PMC

Correlation of BICC1 expression with immune checkpoint expression in PC. A The correlation of BICC1 expression with PDCD1 expression. B The correlation of BICC1 expression with PDCD2 expression. C The correlation of BICC1 expression with CD247 expression. D The correlation of BICC1 expression with CTLA-4 expression. E The correlation of BICC1 expression with HAVCR2 expression. F The correlation of BICC1 expression with LAG3 expression. G The correlation of BICC1 expression with PDCD1LG2 expression. H The correlation of BICC1 expression with TIGIT expression. PC: Pancreatic cancer

Pkd2 was one of the top 15 M6 genes that was most strongly connected with Bicc1 (A). The connection strengths (TOMs, see Methods) are labeled for each Bicc1 gene connection. Pkd2 was the only gene identified from microarray analysis whose expression was significantly (FDR <0.05) correlated with Bicc1 knock down (B). Bicc1 and Pkd2 expression were correlated in osteoblasts isolated from individual Bicc1^+/jcpk and Bicc1^+/+ mice of both sexes (C). Bicc1 and Pkd2 expression were correlated in the HMDP bone samples (D). Causality modeling predicted that Bicc1 is upstream of Pkd2 (E).

The Bicc1 KH domains are required for the function of Bicc1 in embryonic patterning. (A) Validated Bicc1 antisense phosphorothioate oligonucleotide (oligo 9463) was injected into oocytes and the oocytes matured overnight. Matured oocytes were treated with vital dyes, transferred to an ovulating host female and the laid eggs from manipulated oocytes were fertilized. (B-D) Phenotypes of control and sibling experimental Xenopus embryos. Summary presented in G. (B) Control embryos (stage 22). (C) Stage 22 embryos depleted of maternal bicc1 mRNA. The maternal knockout embryos develop with expanded dorsal-anterior structures (DAI 7). (D) The defects from depleting embryos of bicc1 were rescued by wild-type bicc1 mRNA. Embryos depleted of maternal bicc1 mRNA were injected at the vegetal pole with wild-type HA-bicc1 mRNA (20 pg). (E) The defects from depleting embryos of bicc1 were not rescued by KH1-2-3 GDDG bicc1 mRNA. Embryos depleted of maternal bicc1 mRNA were injected at the vegetal pole with KH1-2-3 GDDG bicc1 mRNA (20 pg). (F) The wild-type and KH1-2-3 GDDG Bicc1 proteins were expressed at comparable levels. Proteins from maternal knockout embryos injected with the different mRNAs were analyzed by immunoblotting and probing with an HA antibody. (G) Summary of the phenotypes from control, antisense oligo-injected host-transfer embryos and antisense oligo-injected host-transfer embryos co-injected with either mRNA encoding wild-type HA-Bicc1 or mRNA encoding HA-Bicc1 KH1,2,3 GDDG. The (+) samples received an injection of HA-bicc1 mRNA or HA-bicc1 KH1,2,3 GDDG mRNA whereas the (−) samples did not.

A Absence of left LPM pitx2 expression in bicc1 morphants, unilaterally injected on the left, was rescued by parallel knockdown of dand5. Specificity of TBMO or SBMO was shown by co-injecting rescue mRNAs, i.e., mouse bicc1 or Xenopus bicc1, respectively. Note dand5 knockdown on the right efficiently induced pitx2 expression, as published. B Loss of dand5 mRNA at post-flow stages (st. 20) following left- and right-sided bicc1 SBMO injections. Controls (co) showed wt expression. dand5 expression was restored by co-injecting bicc1 rescue mRNA. Note enhanced dand5 staining in rescued specimens. C Quantification of dand5 expression after knockdown of bicc1. The effect was observed in the left and right sLRO cells. D, E Downregulation of nodal1 in bicc1 morphants. D Quantification of results. E Wt specimens show bilateral nodal1 mRNA. Left or right bicc1 SBMO injections reduced nodal1, which was restored by adding rescue bicc1 mRNA. MO pmol/embryo: bicc1 SBMO (L and S, each 1); bicc1 TBMO, (L and S, each 1); dand5 TPMO (0.5). Asterisks in B and E mark the injected side. Numbers (n) in A, C, and D represent analyzed specimens from more than three independent experiments. Statistical analyses were done with one-sided Pearson’s chi-square test, which was adjusted for multiple comparisons by Bonferroni (B) or Bonferroni–Holm (C, D). n.s. not significant p > 0.05; * significant, p < 0.05; ** highly significant p < 0.01; ***, very highly significant p < 0.001. p values and listing of individual experiments can be found in the source data file. st., stage. Scale bars in B and E represent 100 µm. st. stage, a anterior, l left, r right, p posterior.