Journal: bioRxiv

Article Title: Cargo-Adaptor Cooperation Programs Retromer Coat Architecture

doi: 10.64898/2026.05.05.722683

Figure Lengend Snippet: ( a ) Independent HeLa S3 knockout (KO) cloned cell lines for SNX 3 (clone 1 and 2), SNX12 (clone 1 and 5), and for SNX3 plus SNX12 (3/12) were lysed and subjected to SDS-PAGE followed by immunoblotting with antibodies recognizing SNX3, SNX12, or -actin or GAPDH (as a loading controls). ( b ) Parental HeLa S3 cells (P) and cell clones knocked out for SNX3, SNX12, and SNX3 plus SNX12 (3/12) were infected with wild-type HPV16 PsV. At 48 hpi, cells were analyzed by flow cytometry to quantify HcRed reporter expression. The mean fluorescence intensity (MFI) of parental cells was set to 100%, and data points show the results from four independent cell lines of each genotype (as a percentage of the MFI of infected parental cells), with the means shown by the horizontal lines. Statistical significance was assessed by one-way ANOVA. Significance values: *, p<0.05; ****, p < 0.0001. ( c ) HeLa S3 cells were transfected with small interfering RNA (siRNA) targeting SNX12 or with control scrambled siRNA. 24 hours after transfection, cells were lysed and subjected to SDS-PAGE followed by immunoblotting with antibodies recognizing SNX12, VPS35, or b-actin. ( d ) SNX12 knock down cells as in panel c were mock-infected or infected with wild-type HPV16 L2-3xFLAG PsV 24 hours after transfection with control siRNA (siCont) or siRNA targeting SNX12. At 48 hpi, cells were analyzed by flow cytometry to quantify HcRed expression. The left panel shows a representative histogram, and the right panel shows the quantified infectivity for four independent experiments, expressed as a percentage of MFI of siCont infected cells, with error bars showing standard deviation. Significance values: *, p<0.05; ****, p < 0.0001

Article Snippet: HeLa S3 cells were obtained from the American Type Culture Collection (ATCC), and HEK 293TT cells were gift from Christopher Buck (NIH).

Techniques: Knock-Out, Clone Assay, SDS Page, Western Blot, Infection, Flow Cytometry, Expressing, Fluorescence, Transfection, Small Interfering RNA, Control, Knockdown, Standard Deviation

Journal: bioRxiv

Article Title: Cargo-Adaptor Cooperation Programs Retromer Coat Architecture

doi: 10.64898/2026.05.05.722683

Figure Lengend Snippet: ( a , b ) Control- or SNX12 siRNA-treated HeLa S3 cells were mock-infected or infected with HPV16 L2-3xFLAG for 8 h and subjected to PLA assay using antibodies recognizing FLAG and either SNX12 ( a ) or VPS35 ( b ), as indicated. PLA signals are shown in green, and nuclei were stained with DAPI (blue). PLA signals in singles cells were quantified and plotted in the right panels. ( c and d ) Cells were mock-infected or infected as above for 8 h ( c ) or 16 h ( d ) and subjected to the PLA assay using antibodies recognizing L1 and EEA1. Results are displayed as in panel (a). In all panels, statistical significance was assessed by two-way ANOVA. Significance values: ns, not significant; ***, p < 0.001; and ****, p < 0.0001.

Article Snippet: HeLa S3 cells were obtained from the American Type Culture Collection (ATCC), and HEK 293TT cells were gift from Christopher Buck (NIH).

Techniques: Control, Infection, Staining

Journal: Scientific Reports

Article Title: The C16orf87 protein is a subunit of the MIER corepressor complex controlling embryonic development and cell migration

doi: 10.1038/s41598-026-50740-7

Figure Lengend Snippet: C16orf87 partially mediates HDAC1 and MIER1 protein interactions. ( A ) C16orf87 interacts with the HDAC and MIER proteins. Volcano plot of the IP-MS experiment showing identified proteins interacting with the Flag-C16orf87 protein in HeLa cells. An adjusted P -value cut-off of 0.05 and a log2 fold change cut-off of 2 were used. Data are shown from a biological triplicate experiment. ( B ) Lack of C16orf87 does not change HDAC and MIER protein accumulation. Soluble Panc-01 WT (WT) and Panc-01 KO (KO) whole-cell lysates were analyzed by WB with the indicated antibodies. ( C ) C16orf87 partially mediates HDAC1 and MIER1 interaction. Co-immunoprecipitation of Flag-HDAC1 from siRNA (siC16 and siScr) and pcDNA3-Flag-HDAC1-transfected HeLa cells. Isolated proteins were analyzed by WB with the indicated antibodies. An arrowhead indicates the migration of the MIER1 protein isoforms, whereas an asterisk indicates the migration of the C16orf87 isoforms. ( D ) HDAC1 interacts weakly with C16orf87 in vitro. GST (as a control) and GST-HDAC1 pull-down with bacterially purified 8 × His-tagged C16orf87(Wt, 5 × C > A, 1–130, and 5 × C > A/1–130) proteins. An asterisk indicates a degradation product/partially translated GST-HDAC1. Proteins were detected with the anti-His and anti-GST antibodies.

Article Snippet: Human pancreatic cancer cell lines Panc-01 (ATCC, CRL-1469) and MiaPaCa-2 (ATCC, CRL-1420), mouse skeletal muscle cell line C2C12 (ATCC, CRL-1772), and human cervical cancer cell line HeLa S3 (ATCC, CCL-2.2) were used in this study.

Techniques: Protein-Protein interactions, Immunoprecipitation, Transfection, Isolation, Migration, In Vitro, Control, Purification