human cervical epithelial carcinoma cell line hela cell  (ATCC)

Journal: Cell reports

Article Title: Streptococcus pneumoniae Infection Promotes Histone H3 Dephosphorylation by Modulating Host PP1 Phosphatase.

doi: 10.1016/j.celrep.2020.02.116

Figure Lengend Snippet: Figure 4. PP1 Is the Host Phosphatase Mediating H3S10 Dephosphorylation (A) Representative immunoblot images are shown on the left and quantifications on the right. A549 cells pretreated with the PP1 inhibitor Tautomy- cetin, the PP2A inhibitor okadaic acid, or DMSO as the control and infected with R6 (MOI = 50) for 3 h. (B) A549 cells are transfected with PP1 siRNA or control siRNA before 3 h R6 (MOI = 50) infection. (C) A549 cells are transfected with PP1 siRNA or control siRNA before PLY treatment for 30 min. (D) A549 cells are transfected with PP1 siRNA or control siRNA before H2O2 treatment for 1 h. (E) HeLa cells are pretreated with PP1 inhibitor Tautomycetin before LLO treatment for 20 min. (F) HeLa cells are transfected with PP1 siRNA or control siRNA before LLO treatment for 20 min. All quantification graphs are of at least three in- dependent experiments. All quantifications in graphs show the means ± SD, and statistical sig- nificance was calculated by one-way ANOVA method (Turkey post hoc test). **p < 0.01, ***p < 0.001. See also Figure S4.

Article Snippet: The human cervical carcinoma epithelial cell line HeLa (ATCC CCL-2) cells and human colon carcinoma cell line CaCO2 (ATCC HTB-37) cells were cultured in MEM culture medium supplemented with 1% glutamine, 1 mM sodium pyruvate (GIBCO), 0.1 mM nonessential amino acid solution (GIBCO), and 10% (HeLa) or 20% (CaCO2) FCS.

Techniques: De-Phosphorylation Assay, Western Blot, Control, Infection, Transfection

Journal: Cell reports

Article Title: Streptococcus pneumoniae Infection Promotes Histone H3 Dephosphorylation by Modulating Host PP1 Phosphatase.

doi: 10.1016/j.celrep.2020.02.116

Figure Lengend Snippet: Figure 5. Bacterial Infection Induces Dephosphorylation of PP1 (A) A549 cells were infected for 3 h with WT and the indicated mutants of S. pneumonia strain TIGR4 (MOI = 25). A representative immunoblot (left) and a quantification (right) of four independent experi- ments are shown. Anti-phospho-PP1a (Thr320) antibody (Cell Signaling, 2581s) recognizes all PP1 isoforms (Shimada et al., 2010). The PP1 T320ph levels are normalized to PP1a and to the unin- fected control condition. S. pneumonia. Error bars represent SD, and statistical significance was calculated by one-way ANOVA method (Dunnett’s post hoc test, uninfected as control group). **p < 0.01. (B) The correlation of H3S10ph levels and PP1 T320ph levels of infected cells from four indepen- dent experiments is calculated by the nonpara- metric Spearman’s correlation coefficient method (C) Immunofluorescence of PP1 T320ph in A549 cells under uninfected, 3-h WT TIGR4 infection, 3-h DplyDspxB double mutant infection, and 3-h WT infection at MOI = 25 with 3-h Tautomycetin pretreatment conditions. Size bars represent 50 mm. Quantification of nuclear PP1 T320ph fluorescence intensity from at least two indepen- dent experiments, for more than 300 cells counted for each condition. (D) HeLa cells were infected for 3 h with the WT Listeria (EGD strain) and its hly mutant at MOI = 50. A representative immunoblot of three independent experiments is shown. See also Figure S5.

Article Snippet: The human cervical carcinoma epithelial cell line HeLa (ATCC CCL-2) cells and human colon carcinoma cell line CaCO2 (ATCC HTB-37) cells were cultured in MEM culture medium supplemented with 1% glutamine, 1 mM sodium pyruvate (GIBCO), 0.1 mM nonessential amino acid solution (GIBCO), and 10% (HeLa) or 20% (CaCO2) FCS.

Techniques: Infection, De-Phosphorylation Assay, Western Blot, Control, Mutagenesis

Journal: Journal of cell science

Article Title: 3D correlative electron microscopy reveals continuity of Brucella -containing vacuoles with the endoplasmic reticulum.

doi: 10.1242/jcs.210799

Figure Lengend Snippet: Fig. 5. rBCVs undergo limited fusion. (A) Confocal microscope image of HeLa cell

Article Snippet: Mammalian cell lines and plasmids Human cervical carcinoma epithelial cell line (Hela) (ATCC, CCL-2) was maintained in Dulbecco Modified Eagle Medium (DMEM) supplemented with 10% fetal calf serum (FCS).

Techniques: Microscopy