Figure S3 A). (B) Assessment of the IQCH/calmodulin interaction by proximity ligation assay (PLA). Analysis of the physical proximity of IQCH and calmodulin (CaM) relative to the biological negative control (DNAI2-SPAG6, two proteins from different axonemal complexes) and positive control (DNAI2-DNAH17, two proteins from the same axonemal complex). (C) 3D structures of IQ motifs bound to calmodulin in the calcium free status. Side chains of the residues involved in IQ-CaM interactions are shown as sticks and colored blue or orange for polar and hydrophobic contacts, respectively. Structural and sequence alignments of the IQ motifs from IQCH, IQCG, and Myosin V are shown. " width="100%" height="100%">
Journal: iScience
Article Title: Identification of IQCH as a calmodulin-associated protein required for sperm motility in humans
doi: 10.1016/j.isci.2023.107354
Figure Lengend Snippet: Association of IQCH with calmodulin in flagella and its requirement for sperm capacitation (A) Western blot analysis of protein tyrosine phosphorylation following in vitro capacitation of spermatozoa from the patient and a control individual, at 37°C for 3 h. The data presented for the control individual are representative of the profiles obtained with semen samples from five control individuals displaying normal semen parameters following WHO criteria ( Figure S3 A). (B) Assessment of the IQCH/calmodulin interaction by proximity ligation assay (PLA). Analysis of the physical proximity of IQCH and calmodulin (CaM) relative to the biological negative control (DNAI2-SPAG6, two proteins from different axonemal complexes) and positive control (DNAI2-DNAH17, two proteins from the same axonemal complex). (C) 3D structures of IQ motifs bound to calmodulin in the calcium free status. Side chains of the residues involved in IQ-CaM interactions are shown as sticks and colored blue or orange for polar and hydrophobic contacts, respectively. Structural and sequence alignments of the IQ motifs from IQCH, IQCG, and Myosin V are shown.
Article Snippet: In addition to the experiment, a number of experimental controls were performed: (I) a technical negative control without primary antibody; (II) a biological negative control with non-interacting proteins, such as DNAI2 (mouse monoclonal, Abnova H00064446-M01, 1/320 dilution) and SPAG6 (rabbit polyclonal, Sigma HPA038440, 1/200 dilution); (III) a biological positive control with interacting proteins, such as DNAI2 (mouse monoclonal, Abnova H00064446-M01, 1/320) and DNAH17 (rabbit polyclonal Sigma HPA024354, 1/320 dilution); and (IV) the testing condition with IQCH (rabbit polyclonal, Sigma HPA040845 antibody; 1/100 dilution) and calmodulin (mouse monoclonal, Upstate 05-173; 1/100 dilution).
Techniques: Western Blot, In Vitro, Proximity Ligation Assay, Negative Control, Positive Control, Sequencing
Figure S3 A). (B) Assessment of the IQCH/calmodulin interaction by proximity ligation assay (PLA). Analysis of the physical proximity of IQCH and calmodulin (CaM) relative to the biological negative control 
