Journal: Reproduction (Cambridge, England)

Article Title: Chromatin remodelers HELLS, WDHD1 and BAZ1A are dynamically expressed during mouse spermatogenesis

doi: 10.1530/REP-22-0240

Figure Lengend Snippet: Summary of spermatogenic expression pattern of the proteins analyzed in this study. The expression of HELLS (purple), WDHD1 (green), BAZ1A (red), SCML1 (orange) and SCML2 (blue) in different spermatogenic cell types. Solid line, high expression level. Dashed line, low expression level. Sex body (sphere in pachytene spermatocytes) in stage II–V pachytene spermatocytes stains positively for SCML2 (blue) and HELLS (purple). In stages VI–X ,SCML2 (blue), HELLS (purple) and BAZ1A (red) colocalize in the sex body in WT mouse testis.

Article Snippet: Briefly, following rehydration and antigen retrieval, quenching, permeabilization and blocking (2% normal donkey serum (NDS) + 2% bovine serum albumin (BSA) + 0.1% Triton X-100 in PBS), the samples were incubated overnight at +4°C with following primary antibodies: HELLS (1:500, Millipore, ABD41), WDHD1 (1:500, A301-141A-T), phospho-Histone H2A.X (Ser139) (1:500, Millipore, 05-636), HP1β (1:25; Millipore, MAB3348), BAZ1A (1:500, Sigma, HPA002730), SCML1 (1:100, Santa Cruz, sc-135622) and SCML2 (DSHB Hybridoma Product PCRP-SCML2-1D12 that was deposited to the DSHB by Common Fund – Protein Capture Reagents Program).

Techniques: Expressing

Journal: Journal of proteome research

Article Title: In vivo proximity labeling for the detection of protein-protein and protein-RNA interactions.

doi: 10.1021/pr500196b

Figure Lengend Snippet: Figure 2. Self- and trans-labeling by IPL in the PRC2 complex. (A) Schematic depiction of self-labeling reactions (left) and trans-labeling reaction (right) in the context of the PRC2 complex used for this proof of concept. (B) 293T-REx cells expressing the N-terminal fusions mSA−EZH2 or GAL4−EZH2 (negative control) were subjected to IPL with the indicated concentrations of bio-ASA. Biotinylated proteins were recovered by streptavidin pull-down and revealed by western blots with EZH2 and EED antibodies. (C) IPL of GAL4−EZH2 (control) and mSA−EZH2 followed by IP for PRC2 components (EZH2, EED, and SUZ12) as well as a PRC1-associated factor (SCML2) as an additional control. Western blots for EZH2, EED, and SCML2 (left) and biotin (right) are shown.

Article Snippet: Antibodies against EZH2, EED, and SCML2 were kindly provided by the Reinberg laboratory.20,21 Antibodies against biotin (Bethyl laboratories, TX), SNRNP70 (Santa Cruz Biotech, TX), and SUZ12 (Cell Signaling Technology, MA) were obtained from their respective vendors.

Techniques: Labeling, Expressing, Negative Control, Western Blot, Control

Journal: The EMBO Journal

Article Title: Speedy A governs non-homologous XY chromosome desynapsis as a unique prerequisite for XY loop-axis organization

doi: 10.1038/s44318-025-00528-8

Figure Lengend Snippet: ( A ) X-Y axis length measurements in late pachytene (RA13) Sync-Spdya fl/fl (a) and Sync-Spdya cKO (b) spermatocytes. Green and blue dashed lines show X and Y chromosome axes, respectively. (c–e) Measured lengths of the X axis (c), Y axis (d), and the calculated Y/X axis length ratio (e) in late pachytene Sync-Spdya fl/fl cells and Sync-Spdya cKO cells with full Y-X NH synapsis . Each dot represents a single nucleus. ( B ) Immuno-FISH staining of late pachytene Sync-Spdya fl/fl (a–a″) and Sync-Spdya cKO (b–b″) spermatocytes for γH2AX (light blue), Chr Y (green) and SYCP3 (red). The white dashed line-enclosed area indicates Y chromatin signals located outside the γH2AX area. Scale bars, 2 µm. (c) Percentage of Y chromatin loop signals outside γH2AX area in Sync-Spdya cKO spermatocytes, based on the fluorescence intensity measurements of Y loops inside and outside the γH2AX domain. ( C ) Immuno-FISH staining of late pachytene Sync-Spdya fl/fl (a–a″) and Sync-Spdya cKO (b–b″) spermatocytes for SCML2 (light blue), Chr Y (green) and SYCP3 (red). The white dashed line-enclosed area indicates Y chromatin signals located outside the SCML2 area. Scale bars, 2 µm. (c) Percentage of Y chromatin loop signals outside SCML2 area in Sync-Spdya cKO spermatocytes. ( D ) Sync-Spdya fl/fl (a) and Sync-Spdya cKO (b) spermatocytes immunostained for FK2 (green), SYCP3 (red) and ACA (light blue) showing the absence of FK2 signals in the full Y-X NH synapsis region (dashed circle in (b)) of X-Y chromosomes in Sync-Spdya cKO spermatocytes. Scale bars, 2 µm. (c) Percentage of late pachytene Sync-Spdya cKO spermatocytes lacking FK2 signals at the full Y-X NH synapsis region. ( E ) Illustration of a proposed mechanism underlying SpdyA-mediated X-Y loop-axis organization at late pachynema. Data information: All values are presented as the mean ± SD. “ n ” represents the total number of pachytene spermatocytes scored per genotype: from two mice in ( A – C ), and from three mice in ( D ). Statistical analyses were performed using Mann–Whitney test, and both exact P values in ( A -d) and ( A- e) ˂ 0.000000000000001. .

Article Snippet: The germline-specific Polycomb protein SCML2 (Sex comb on midleg-like 2) is recruited to X-Y chromatin at the early-to-mid-pachytene transition, suppressing the ubiquitination of H2A Lysine 119 (K119) on the X-Y chromosomes (Hasegawa et al, ; Luo et al, ).

Techniques: Staining, Fluorescence, MANN-WHITNEY