Journal: Human molecular genetics

Article Title: Biallelic missense variants in ZBTB11 can cause intellectual disability in humans.

doi: 10.1093/hmg/ddy220

Figure Lengend Snippet: Figure2. In-silico 3D models of the two missense variants are shown. A. ZBTB11 WT C2H2-type zinc finger

Article Snippet: In addition, anti-ZBTB11 Antibody (#HPA015328; Atlas Antibodies, Stockholm, Sweden) with Goat Anti-Rabbit IgG H&L (FITC) (#ab6717; Abcam, Cambridge, USA), secondary antibodies were used to assess expression of endogenous ZBTB11 in HEK293 cell lines.

Techniques: In Silico

Journal: Human molecular genetics

Article Title: Biallelic missense variants in ZBTB11 can cause intellectual disability in humans.

doi: 10.1093/hmg/ddy220

Figure Lengend Snippet: Figure3. Cellular Localization of wild-type (WT) recombinant ZBTB11, and the mutants (H729Y and H880Q)

Article Snippet: In addition, anti-ZBTB11 Antibody (#HPA015328; Atlas Antibodies, Stockholm, Sweden) with Goat Anti-Rabbit IgG H&L (FITC) (#ab6717; Abcam, Cambridge, USA), secondary antibodies were used to assess expression of endogenous ZBTB11 in HEK293 cell lines.

Techniques: Recombinant

Journal: bioRxiv

Article Title: Targeting transcription factors through an IMiD independent zinc finger domain

doi: 10.1101/2024.01.03.574032

Figure Lengend Snippet: A) 293T cells were stably transfected with pFUW-SALL4B wild type (SALL4B WT ), or pFUW-SALL4B with deletion of ZFC4 (SALL4B -ZFC4 ). Cells were treated with SH6, Lenalidomide (Len), or DMSO for 24 hours before harvested for western blot analysis. SH6 mediated degradation of SALL4B was abolished in the absence of ZFC4, while Lenalidomide did not affect SALL4B levels in SALL4B -ZFC4 or SALL4B WT cells. B) Degrome analysis revealed two additional ZFC4 containing proteins. A volcano plot from the mass spectrometry data demonstrates the change of the cellular proteins from the SH6 vs DMSO treatment. Down regulated proteins with twofold change and p-value of 0.05 were highlighted in red. In brief, SNU398 was grown treated with SH6 (50uM) or DMSO for 24hr. C) Sequence alignment of the SALL4 ZFC4 domain with ZBTB7A and ZBTB11. α-helices (α) and β-strands (β) are indicated according to data on crystal structures of SALL4 ZFC4. The colour scheme used for residue conservation is based on side chain properties. Blue=hydrophobic (A, I, L, M, F, W, V); Red=positive charge (K, R); Magenta= negative (D, E); Green= polar (S, T, N, Q); Pink= cysteine; Orange= glycine; Yellow= proline; Cyan= aromatic (Y, H). D) Expression levels of ZBTB7A and ZBTB11 in DMSO/ SH6 treated samples analysed by mass spectrometry. Data represent the mean ± s.d. (n=3). E) SNU398 cells were treated with DMSO, Pomalidomide, or SH6 for 24 hours, harvested, and analysed by western blot. Blots were probed with ZBTB7A ( E(i) ) or ZBTB11 ( E(ii) ) antibodies. F) The ZBTB7A crystal structure (PDB 7N5W; Green) was overlaid with that of SALL4 ZFC4 (Red) and analysed with PyMol. Overlapped regions are shown.

Article Snippet: After blocking in Blocking One (Nacalai Tesque), the membrane was probed with primary antibodies to SALL4 (Santa Cruz Biotechnology, sc-101147), β-Actin (Santa Cruz Biotechnology, sc-47778), Cul4A (#2699) Cell Signalling Technology, CRBN (D8H3S, #71810) Cell Signalling Technology, LRF (13E9, 14-3309) from eBioscience, Armenian hamster IgG-HRP (sc-2789) from Santa Cruz, and ZBTB11 antibody (25215-1-AP) from Proteintech, and incubated overnight at 4 °C.

Techniques: Stable Transfection, Transfection, Western Blot, Mass Spectrometry, Sequencing, Residue, Expressing