Journal: Cell Host & Microbe

Article Title: Determinants of Spike infectivity, processing, and neutralization in SARS-CoV-2 Omicron subvariants BA.1 and BA.2

doi: 10.1016/j.chom.2022.07.006

Figure Lengend Snippet:

Article Snippet: Thereafter, samples were washed with PBS and incubated for 2 h at 4°C with primary antibody (anti-V5(Mouse) (1:1,000, Cell Signalling, #80076S)) diluted in PBS.

Techniques: Recombinant, Mutagenesis, Binding Assay, Plasmid Preparation, Software, Microscopy, Modification

Journal: Nature Communications

Article Title: Nuclear pore protein NUP210 depletion suppresses metastasis through heterochromatin-mediated disruption of tumor cell mechanical response

doi: 10.1038/s41467-021-27451-w

Figure Lengend Snippet: a Coimmunoprecipitation of myc-tagged NUP210 with SUN1 and SUN2. b Colocalization of SUN2 and myc-tagged NUP210. Scale bar = 5 μm. c Distribution of SUN1 and SUN2 in Nup210 KO (KO-N13) 4T1 cells. Scale bar = 10 μm. d Quantification of SUN2 and SUN1 intensity in Nup210 KO 4T1 cells. Mann–Whitney U test, error bar represents median with an interquartile range. ‘ n ’ equals the number of cells analyzed. e Distribution of Lamin B1 and Lamin A/C in Nup210 KO 4T1 cells. MIP maximum intensity projection. Scale bar = 5 μm. f Quantification of Lamin B1 and Lamin A/C intensity in Nup210 KO 4T1 cells. Mann–Whitney U test, error bar represents median with an interquartile range. ‘ n ’ equals the number of cells analyzed. g 3D reconstruction of Lamin B1, Lamin A/C, and DAPI (heterochromatin foci) distribution in Nup210 KO 4T1 cells. Scale bar = 2 μm. h Subcellular fractionation of LINC complex proteins and mechanosensitive MRTF-A and YAP in Nup210 KO 4T1 cells. i Co-IP of H3.1/3.2 with LINC complex proteins SUN2, BRD4 short isoform (BRD4-SF), and RRP1B in 4T1 cells. j Co-IP of V5-tagged BRD4-SF with NUP210 in 4T1 cells. k Co-IP of H3.1/3.2 and SUN2 in human MCF7 and MDA-MB-231 cell line. l Co-IP of H3.1/3.2 with NUP210 and SUN2 in JQ1-treated 4T1 cells. m Distribution of H3.1/3.2 and H3K27me3 in 4T1 cells treated with bromodomain inhibitor JQ1. Scale bar = 5 μm. n Quantification of H3.1/3.2 and H3K27me3 intensity in JQ1-treated 4T1 cells. Mann–Whitney U test, error bar represents median with an interquartile range. ‘ n ’ equals the number of cells analyzed. o Live-cell tracking of nuclear size in DMSO control ( n = 122) and JQ1-treated 4T1 cells ( n = 127). p qRT-PCR of NUP210-regulated genes in JQ1-treated 4T1 cells. Two-tailed t test, mean ± s.e.m. n = 3 biological replicates. q qRT-PCR of NUP210-regulated genes in BRD4-SF knockdown 4T1 cells. Two-tailed t test, mean ± s.e.m. n = 3.

Article Snippet: Primary antibodies and their dilutions were as follows: NUP210 (1:500; Bethyl Laboratories), β-actin (1:10,000; Abcam), Lamin B1 (1:5000; Abcam), Lamin A/C (1:1000; Abcam), Myc-Tag (1:1000; Cell Signaling Technology), rabbit V5-Tag (1:1000; Cell Signaling Technology), mouse V5-Tag (1:1000; Cell Signaling Technology), RRP1B (1:1000; Millipore-Sigma), SUN1 (1:500; Abcam), SUN2 (1:1000; Abcam), FAK (1:5000; Abcam), p-FAK(Y397) (1:5000; Abcam), p-FAK (Y397) (1:5000; Thermo Fisher Scientific), CCL2 (1:1000; Proteintech), H3.1/3.2 (1:1000; Active Motif), H3K27me3 (1:5000; Cell Signaling Technology), H3K9me3 (1:5000; Abcam), SUV39H1 (1:1000, Cell Signaling Technology), EZH2 (1:1000; Cell Signaling Technology), and SUZ12 (1:1000; Cell Signaling Technology).

Techniques: MANN-WHITNEY, Fractionation, Co-Immunoprecipitation Assay, Cell Tracking Assay, Control, Quantitative RT-PCR, Two Tailed Test, Knockdown