spib  (Bio-Rad)

Journal: Genes

Article Title: NKX6-3 in B-Cell Progenitor Differentiation and Leukemia

doi: 10.3390/genes16101199

Figure Lengend Snippet: Regulatory connections of NKX6-3 with TALE-class homeobox genes IRX1 and MEIS1 and ETS-TF SPIB. ( A ) SiRNA-mediated knockdown of NKX6-3 in RCH-ACV was confirmed by RQ-PCR (left) and Western blot analysis (right). ( B ) RQ-PCR analysis of RCH-ACV after knockdown of NKX6-3 (left) and IRX1 (right). ( C ) RQ-PCR analysis of RCH-ACV after forced expression of MEIS1 (left) and of SEM after forced expression of NKX6-3 (right). ( D ) RQ-PCR analysis of RCH-ACV after knockdown of NKX6-3 (left) and forced expression of SPIB (right). The data were generated in triplicate. Statistical significance was assessed by t-test (two-tailed), and the calculated p -values are indicated by asterisks (** p < 0.01, *** p < 0.001, n.s. not significant).

Article Snippet: The following primer sets were used: CD109 (Hs00370347_m1), GP5 (Hs03027242_s1), IRX1 (Hs00411782_m1), MEIS1 (Hs01017441_m1), MPP7 (Hs00399584_m1), NKX6-3 (Hs04190028_g1), PBX1 (Hs00231228_m1), SMAD4 (Hs00929647-m1), SPIB (Hs01548149_m1), TCF3 (Hs01012685_m1), TBP (Hs00427620_m1) and TGFBR2 (Hs00234253_m1).

Techniques: Knockdown, Western Blot, Expressing, Generated, Two Tailed Test

Journal: Cell reports

Article Title: Regulatory network analysis of Dclk1 gene expression reveals a tuft cell-ILC2 axis that inhibits pancreatic tumor progression

doi: 10.1016/j.celrep.2025.115734

Figure Lengend Snippet: (A) UMAP showing VIPER-inferred differential protein activity of IL-4RA in Dclk1 + cells. (B and C) Flow cytometry analysis for IL-13 expression in immune cells (CD45) isolated from normal pancreas and pancreas from Mist1-Kras mice and related quantification. (D and E) Flow cytometry analysis for EpCAM and ZsGreen of organoids from pancreas of Kras-Dclk1-DTR-ZsGreen mice in the presence of vehicle or IL-13 and related quantification. (F) RT-qPCR for the expression of Dclk1 . (G) RT-qPCR for the expression of Il13 in different immune cell populations in pancreas from Mist1-Kras mice: CD45 + immune cells, CD4 + T cells, γδ T cells, and ILC2s (Lin — CD45 + CD127 + CD90 + KLRG1 + ). (H and I) Flow cytometry analysis for ILC2s in normal and Mist1-Kras mice and related quantification. (J and K) ZsGreen expression in organoids from Kras-Dclk1-DTR-ZsGreen pancreas cultured with or without ILC2s, with quantification. (L) RT-qPCR for the expression of Dclk1 in organoids. (M) IF for ZsGreen of pancreas from Mist1-Kras-Dclk1-DTR-ZsGreen mice treated with control IGG or anti-IL-13 blocking antibody for 8 weeks. (N and O) Flow cytometry analysis for EpCAM and ZsGreen and related quantification. (P–R) RT-qPCR for the expression of Dclk1 , Cd24a , and Spib . Scale bars: 100 μm. Means ± SD. Statistical significance was evaluated by Student’s t test; * p ≤ 0.05, ** p ≤ 0.01, and **** p ≤ 0.001; n.s., not significant.

Article Snippet: For experiments involving viral infections, single cell suspensions in Matrigel were added with Spib overexpressing lentivirus (VectorBuilder) or Adeno-CMVCre (University of Iowa) at 500 PFU/cell and sub-sequently grown as described above.

Techniques: Activity Assay, Flow Cytometry, Expressing, Isolation, Quantitative RT-PCR, Cell Culture, Control, Blocking Assay