immunoculttm human th1 differentiation supplement  (STEMCELL Technologies Inc)

Journal: Respiratory Research

Article Title: SERPINB10 contributes to asthma by inhibiting the apoptosis of allergenic Th2 cells

doi: 10.1186/s12931-021-01757-1

Figure Lengend Snippet: Serpinb10-knockdown mice had diminished numbers of Th2 cells after challenge. a Gating strategy to identify Th cells and their apoptosis in mouse lungs. Live cells can be separated in a CD45 + and a CD45 − cell population. Within the CD45 + cell population, gate for CD4 T cells (CD3 + CD4 + ). Within the CD4 T cells, gate for Th1 cells (T-bet + ) and Th2 cells (GATA-3 + ). Within the Th1 cells and Th2 cells, gate for apoptosis cells (Active Caspase-3 + ) respectively. b The numbers of Th1 and Th2 cells in mouse lungs. c percentage of caspase-3-positive Th1 and Th2 cells in mouse lungs. Data were combined from three experiments. n = 12–14. *, P < 0.05.

Article Snippet: ImmunoCult Human Th1 Differentiation Supplement (StemCell Technologies) and ImmunoCult Human Th2 Differentiation Supplement (StemCell Technologies) were added to direct the differentiation of Th1 and Th2 cells, respectively.

Techniques: Knockdown

Journal: Respiratory Research

Article Title: SERPINB10 contributes to asthma by inhibiting the apoptosis of allergenic Th2 cells

doi: 10.1186/s12931-021-01757-1

Figure Lengend Snippet: Expression level of SERPINB10 in human Th cells. a flow cytometric plots for Th1 or Th2 polarized CD4 T cells from patients with asthma after intracellular staining for IFN-γ or IL-4. b the transcript level of IFNG, T-bet, IL-4 and GATA-3 in polarized Th1 and Th2 cells was measured using quantitative PCR. c cytokine levels in culture supernatant of polarized Th1 or Th2 cells were measured by LEGENDplex human Th cytokine panel. d SERPINB10 transcript level (fold change over naïve CD4 T cells) in Th1 or Th2 cells after no stimulation or stimulation with anti-CD3 or cytokine (IFN-γ for Th1 and IL-4 for Th2). n = 16 patient samples. Data were means ± SE. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant

Article Snippet: ImmunoCult Human Th1 Differentiation Supplement (StemCell Technologies) and ImmunoCult Human Th2 Differentiation Supplement (StemCell Technologies) were added to direct the differentiation of Th1 and Th2 cells, respectively.

Techniques: Expressing, Staining, Real-time Polymerase Chain Reaction

Journal: Respiratory Research

Article Title: SERPINB10 contributes to asthma by inhibiting the apoptosis of allergenic Th2 cells

doi: 10.1186/s12931-021-01757-1

Figure Lengend Snippet: Knockdown of SERPINB10 expression affects survival of Th2 cells. a representative light micrograph of lentivirus (LV)-transduced CD4 T cells (top and bottom panels are pictures of the same cells in fluorescence mode and normal light mode respectively). b and c flow cytometric data for green fluorescent protein (GFP) + percentage of lentivirus-transduced CD4 T cells. d SERPINB10 transcript level in lentivirus-transduced CD4 T cells (fold change compared with ACTB housekeeping gene). e percentage of IFN-γ + or IL-4 + polarized CD4 cells upon completion of Th1 and Th2 polarization. f percentage of IFN-γ + or IL-4 + polarized CD4 cells after completion of polarization and rested for another 8 days. g flow cytometric histogram for negative control (solid histogram) or SERPINB10-knockdown (line) Th1 and Th2 cells after staining with AnnexinV. Percentage of AnnexinV-positive cells were shown in histograms. Data were pooled from three independent experiment; n = 4 wells per group for each independent experiment. Data were mean ± SE. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant

Article Snippet: ImmunoCult Human Th1 Differentiation Supplement (StemCell Technologies) and ImmunoCult Human Th2 Differentiation Supplement (StemCell Technologies) were added to direct the differentiation of Th1 and Th2 cells, respectively.

Techniques: Knockdown, Expressing, Fluorescence, Negative Control, Staining