Journal: bioRxiv

Article Title: A validated panel of commercial antibodies for reliable detection of FET proteins

doi: 10.1101/2025.11.19.689317

Figure Lengend Snippet: Epitopes of the different anti-FUS, anti-EWS and anti-TAF15 antibodies have been indicated where they are known. Domains are labelled for their function/make up: low complexity domain (QGSY Rich) RNA recognition motif (RRM) and Zinc Finger domain (ZnF), each flanked by arginine-glycine–glycine (RGG)-rich domains, and a C-terminal PY-Nuclear localisation signal (NLS).

Article Snippet: Furthermore, the Cell Signalling-28409 anti-TAF15 antibody showed strong nuclear staining accompanied by some cytoplasmic staining with a mean nuclear intensity of 89%.

Techniques:

Journal: bioRxiv

Article Title: A validated panel of commercial antibodies for reliable detection of FET proteins

doi: 10.1101/2025.11.19.689317

Figure Lengend Snippet: Western blot validation of seven commercially available anti-FUS antibodies showed a single band at ∼70 kDa corresponding to FUS. All antibodies show a decrease of FUS protein levels in FUS only KD (Lane 2), FUS + EWS KD (Lane 5) and FUS + TAF15 KD (Lane 6). All bands were normalised to GAPDH loading control (37 kDa) and ratios of FET single/double knockdowns were then compared to control and significance determined by performing One-Way ANOVA with post-hoc Tukey’s multiple comparisons test (See Figure S2 for quantifications).

Article Snippet: Furthermore, the Cell Signalling-28409 anti-TAF15 antibody showed strong nuclear staining accompanied by some cytoplasmic staining with a mean nuclear intensity of 89%.

Techniques: Western Blot, Biomarker Discovery, Control

Journal: bioRxiv

Article Title: A validated panel of commercial antibodies for reliable detection of FET proteins

doi: 10.1101/2025.11.19.689317

Figure Lengend Snippet: Western blot validation of four commercially available anti-FUS antibodies showed a single band at ∼70 kDa corresponding to FUS. All antibodies show a decrease of FUS protein levels in FUS only KD (Lane 2), FUS + EWS KD (Lane 5) and FUS+TAF15 KD (Lane 6). However, all four showed additional bands identified as EWS and/or TAF15 as well as those that don’t appear to recognise any FET protein. All bands were normalised to GAPDH loading control (37 kDa) and ratios of FET single/double knockdowns were then compared to control and significance determined by performing One-Way ANOVA with post-hoc Tukey’s multiple comparisons test (See Figure S4 for quantifications).

Article Snippet: Furthermore, the Cell Signalling-28409 anti-TAF15 antibody showed strong nuclear staining accompanied by some cytoplasmic staining with a mean nuclear intensity of 89%.

Techniques: Western Blot, Biomarker Discovery, Control

Journal: bioRxiv

Article Title: A validated panel of commercial antibodies for reliable detection of FET proteins

doi: 10.1101/2025.11.19.689317

Figure Lengend Snippet: Western blot validation of seven commercially available anti-EWS antibodies showed a single band at ∼80 kDa corresponding to EWS. All antibodies show a decrease of EWS protein levels in EWS only KD (Lane 3), FUS + EWS KD (Lane 5) and TAF15 +EWS KD (Lane 7). All bands were normalised to GAPDH loading control (37 kDa) and ratios of FET single/double knockdowns were then compared to control and significance determined by performing One-Way ANOVA with post-hoc Tukey’s multiple comparisons test (See Figure S6 for quantifications).

Article Snippet: Furthermore, the Cell Signalling-28409 anti-TAF15 antibody showed strong nuclear staining accompanied by some cytoplasmic staining with a mean nuclear intensity of 89%.

Techniques: Western Blot, Biomarker Discovery, Control

Journal: bioRxiv

Article Title: A validated panel of commercial antibodies for reliable detection of FET proteins

doi: 10.1101/2025.11.19.689317

Figure Lengend Snippet: Western blot validation of seven commercially available anti-TAF15 antibodies showed a main band at ∼77 kDa corresponding to TAF15. Most antibodies (A, B, D, E, F & G) had an additional lower band determined to be TAF15. All antibodies show a decrease of the main TAF15 band in TAF15 only KD (Lane 4), FUS + TAF15 KD (Lane 6) and EWS + TAF15 KD (Lane 7). The lower band increased in the FUS+EWS KD (Lane 5). All bands were normalised to GAPDH loading control (37 kDa) and ratios of FET single/double knockdowns were then compared to control and significance determined by performing One-Way ANOVA with post-hoc Tukey’s multiple comparisons test (See Figure S8 for quantifications).

Article Snippet: Furthermore, the Cell Signalling-28409 anti-TAF15 antibody showed strong nuclear staining accompanied by some cytoplasmic staining with a mean nuclear intensity of 89%.

Techniques: Western Blot, Biomarker Discovery, Control

Journal: bioRxiv

Article Title: A validated panel of commercial antibodies for reliable detection of FET proteins

doi: 10.1101/2025.11.19.689317

Figure Lengend Snippet: anti-TAF15 antibody immunofluorescence detection of endogenous TAF15 (Green) in primary cortical neurons and wild-type HeLa cells. MAP2 (red) is used as a neuronal marker for primary cortical neurons, while Cell Mask (red) is used to stain the plasma membrane of HeLa cells. Nuclei are labelled with DAPI (blue). n = 3. 40x magnification. Scale bar (primary cortical neurons) = 100μm, Scale bar (HeLa cells)= 20μm. See Figure S10 for quantification graphs.

Article Snippet: Furthermore, the Cell Signalling-28409 anti-TAF15 antibody showed strong nuclear staining accompanied by some cytoplasmic staining with a mean nuclear intensity of 89%.

Techniques: Immunofluorescence, Marker, Staining, Clinical Proteomics, Membrane

Journal: Aging cell

Article Title: TAF15 downregulation contributes to the benefits of physical training on dendritic spines and working memory in aged mice.

doi: 10.1111/acel.14244

Figure Lengend Snippet: FIGURE 5 Physical running downregulates TAF15 expression in the PrL of aged mice. (a, b) A decreased TAF15 expression was identified in aged runners (F2,25 = 46.80; **p < 0.01; n = 9 or 10). The protein levels of TAF15 were normalized to respective Actin levels and expressed as a percentage of adult controls. (c) TAF15 was primarily detected in DAPI-counterstained nuclei. The co-localization was pointed by arrowheads. (d–f) TAF15 was detected in NeuN-labeled neurons (arrowheads), but not Iba1-labeled microglial cells and GFAP-labeled astrocytes. (g–i) Representative confocal images from an adult, aged control, and aged runner to show TAF15-ir nuclei in layers V–VI. (j) A decreased number of TAF15-ir nuclei in the aged runners versus aged controls (F2,25 = 53.68; **p < 0.01). (k) Negative correlations were observed between the numbers of TAF15-ir nuclei (per 5 × 104 μm2) and the spontaneous alternations (left, r = −0.76, p < 0.01) as well as rewarded correct choices (right, r = −0.79, p < 0.01). (l) A negative correlation between TAF15-ir nuclei and total dendritic spines (r = −0.80, p < 0.01). Scale bars = 20 μm (c, d–i).

Article Snippet: In brief, free- floating sections were treated in 0.3% H2O2 in PBS- T for 30 min, followed by a blockade of nonspecific sites with 5% normal serum for 1 h. Sections were incubated in mouse anti- GFP (1:5000; Sigma), rabbit anti- TAF15 (1:1000; Bethyl, IHC- 00094), rabbit anti- TAF15- 309A (1:20,000; Bethyl), mouse anti- NeuN (1:1000; Chemicon, MAB377), mouse anti- Iba1 (1:2000; Synaptic Systems, #234011), and mouse antiGFAP (1:5000; Sigma, cat # 3402) 1–3 days at 4°C.

Techniques: Expressing, Labeling, Control

Journal: Aging cell

Article Title: TAF15 downregulation contributes to the benefits of physical training on dendritic spines and working memory in aged mice.

doi: 10.1111/acel.14244

Figure Lengend Snippet: FIGURE 5 Physical running downregulates TAF15 expression in the PrL of aged mice. (a, b) A decreased TAF15 expression was identified in aged runners (F2,25 = 46.80; **p < 0.01; n = 9 or 10). The protein levels of TAF15 were normalized to respective Actin levels and expressed as a percentage of adult controls. (c) TAF15 was primarily detected in DAPI-counterstained nuclei. The co-localization was pointed by arrowheads. (d–f) TAF15 was detected in NeuN-labeled neurons (arrowheads), but not Iba1-labeled microglial cells and GFAP-labeled astrocytes. (g–i) Representative confocal images from an adult, aged control, and aged runner to show TAF15-ir nuclei in layers V–VI. (j) A decreased number of TAF15-ir nuclei in the aged runners versus aged controls (F2,25 = 53.68; **p < 0.01). (k) Negative correlations were observed between the numbers of TAF15-ir nuclei (per 5 × 104 μm2) and the spontaneous alternations (left, r = −0.76, p < 0.01) as well as rewarded correct choices (right, r = −0.79, p < 0.01). (l) A negative correlation between TAF15-ir nuclei and total dendritic spines (r = −0.80, p < 0.01). Scale bars = 20 μm (c, d–i).

Article Snippet: In brief, free- floating sections were treated in 0.3% H2O2 in PBS- T for 30 min, followed by a blockade of nonspecific sites with 5% normal serum for 1 h. Sections were incubated in mouse anti- GFP (1:5000; Sigma), rabbit anti- TAF15 (1:1000; Bethyl, IHC- 00094), rabbit anti- TAF15- 309A (1:20,000; Bethyl), mouse anti- NeuN (1:1000; Chemicon, MAB377), mouse anti- Iba1 (1:2000; Synaptic Systems, #234011), and mouse antiGFAP (1:5000; Sigma, cat # 3402) 1–3 days at 4°C.

Techniques: Expressing, Labeling, Control