chelex-100 sodium form 50 to 100 mesh  (Millipore)

Journal: Cell Death Discovery

Article Title: Ca 2+ signals are essential for T-cell proliferation, while Zn 2+ signals are necessary for T helper cell 1 differentiation

doi: 10.1038/s41420-024-02104-1

Figure Lengend Snippet: A PBMC were stimulated at the same time with pyrithione and thapsigargin. Protein expression of the transcription factor FoxP3 was examined by western blot 48 h after stimulation and normalized to β-actin. A representative western blot is shown. B In addition, PBMC were preincubated for 15 min in normal or Chelex-treated Zn 2+ -deficient medium and were subsequently stimulated with thapsigargin. FoxP3 was also examined by western blot after 48 h and normalized to β-actin. A representative western blot is shown. C PBMC were stimulated at the same time with pyrithione and thapsigargin. 3 h after stimulation KLF-10 expression was examined by qPCR and normalized to PBDG. D In addition, PBMC were preincubated with the p38 MAPK inhibitor SB202190 (10 µM) for 30 min and then stimulated with pyrithione and thapsigargin. KLF-10 expression was also examined after 3 h by qPCR. The control data shown in ( D ) are also included in the data shown in Fig. 7C. Data are presented as mean + SEM with n = 9 ( A ), n = 6 ( B ), n = 12 ( C ) and n = 7 ( D ) experiments. Statistical significance was determined by one-way ANOVA with Tukey’s multiple comparisons test ( A – C ) or with Sidak’s multiple comparisons test ( D ). Significantly different results ( p < 0.05) have no common identification letter ( A , C ) and * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: To incubate cells in medium without Zn 2+ , the described medium was treated for 1 h with Chelex ® 100 sodium form (Sigma-Aldrich) to chelate all divalent cations.

Techniques: Expressing, Western Blot, Control