Fig 1: HNRNPH1 regulates the alternative splicing of cell cycle-associated genes.A, B Analysis of HNRNPH1 binding sites in the AURKB (A) and ESPL1 (B) pre-mRNA by eCLIP. Read density in reads per million (RPM) is shown for HNRNPH1, IgG, and input. C HNRNPH1 RIP (RNA immunoprecipitation) followed by qRT-PCR analysis of co-purified AURKB (left) and ESPL1 (right) in UV-crosslinked PDGCs. D qRT-PCR analysis of exon-intron and exon-exon junction for AURKB (left) and ESPL1 (right). E Diagram of AURKB minigene splicing with primers used for PCR analysis (top). Wild-type (wt) and mutant (mut) sequences in the intronic region of the minigene are shown (bottom). Red letters represent the mutated nucleotides. F Splicing pattern of the transfected empty control plasmid, wt AURKB minigene, and mut AURKB minigene in HEK293 cells. G Distribution of alternative splicing events detected using the Junction Count (JC) approach. H Differentially spliced events detected by the JC approach. Inclusion level differences (U87 Control vs. HNRNPH1-knockout) on the x-axis and statistical significance (-log10 FDR) on the y-axis. Significant events (FDR < 0.05, |ΔPSI | > 0.05) are highlighted for higher inclusion (orange) and higher exclusion (blue). I Sashimi plot showing the inclusion of UHRF2 exon 9a in U87 HNRNPH1-knockout (blue) compared to U87 HNRNPH1 wild-type (red). J Analysis of UHRF2 exon 9a percent inclusion (PSI%) in U87 HNRNPH1-knockout compared to U87 control. K Diagram of UHRF2 AS (top) and PCR analysis of UHRF2 splicing pattern in G62 PDGCs transfected with siRNA control or siRNA targeting HNRNPH1 (bottom). L qRT-PCR analysis of total UHRF2 (left) and UHRF2 variant 2 (exon 9-exon 9a junction) expression in G62 PDGCs transfected with siRNA control or siRNA targeting HNRNPH1. M Western Blot analysis of UHRF2 and CCND1 expression in G62 PDGCs transfected with siRNA control or siRNA targeting HNRNPH1. Data shown as mean ± SD of three replicates. Data were analyzed by unpaired t-test: **p < 0.01, ***p < 0.001, ****p < 0.0001.