Journal: Materials Today Bio

Article Title: A multifunctional and ROS response CO-gas delivery platform for spinal cord regeneration

doi: 10.1016/j.mtbio.2026.102760

Figure Lengend Snippet: COPH could decreases neuronal apoptosis at 7 d.p.i. (A) Immunofluorescence staining of NeuN (green), C-caspase3 (red) and DAPI (blue) in different groups. Scale bar: 50 μm. (B) Western blotting showing the expression of Bax, Bcl-2 and C-caspase3 in different groups 7 d.p.i. (C) Statistical analysis of C-caspase3 fluorescence intensity in each group. (D–F) The quantitative analysis of Bax, Bcl-2 and C-caspase3 protein expression. ∗∗ represents P < 0.01 vs SCI group. ## represent COPs group vs COPH group respectively p < 0.01, (n = 3). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Primary antibody to C-Caspase3, CD206, IL-1β, HO-1, SOD1, P65 was provided by Affinity Biosciences (USA).

Techniques: Immunofluorescence, Staining, Western Blot, Expressing, Fluorescence

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Uncovering the mechanisms of homologous point acupuncture on knee osteoarthritis through an integrated study of metabolomics and proteomics

doi: 10.3389/fbioe.2026.1791109

Figure Lengend Snippet: Effects of HAP on cartilage autophagy and apoptosis in KOA model rats. Note: (A,C–F) Expression of LC3 I, LC3 II, Beclin1, C-Caspase3 and Bax in cartilage tissue (GAPDH as internal control). (B,G,H) Expression of p62 and Bcl2 in cartilage (GAPDH as internal control). *** p < 0.001, ** p < 0.01, * p < 0.05.

Article Snippet: All the primary antibodies were validated by either the manufacturer or other available publications, listed as follows: HIF1α (Boster, A00013-1), HK2 (Abclonal, A0994), c-caspase3 (Affinity, AF7022), Bax (Wuhan Sanying, 50599-2-Ig), LC3 (CST, 12741T), Beclin1 (Wuhan Sanying, 11306-1-AP), and GAPDH (Wuhan Sanying, 60004-1-Ig) from WUHAN HUAYAN Biotechnology CO., LTD. NLRP3 (Affinity, BF8029), ASC (Affinity, DF6304), caspase1 (Affinity, AF5418), p62 (Wuhan Sanying, 18420-1-AP), Bcl2 (Wuhan Sanying, 60178-1-Ig) from WUHAN Fabre Biotechnology CO., LTD. After washing 5 times for 5 min each with TBST (T-Pro), and a horseradish peroxidase (HRP)-conjugated secondary antibody (Boster, BA1051 and BA1054) was applied for 2 h at room temperature.

Techniques: Expressing, Control

Journal: bioRxiv

Article Title: PTK2 regulates the UPS impairment via p62 phosphorylation in TDP-43 proteinopathy

doi: 10.1101/355446

Figure Lengend Snippet: (A-C) N2a cells were transiently transfected with the control vector (pCMV6-AC-GFP) or TDP-43 expression construct (pCMV6-AC-TDP-43-GFP) for 3 days. Cells overexpressing GFP or TDP-43-GFP were treated with ptk2 -specific siRNA (20 nM) or a PTK2 inhibitor (5 μm) for 24 h. Immunocytochemistry was performed thereafter. (A) Immunocytochemistry was subsequently performed to detect PTK2 phosphorylation (p-PTK2 Y397 ; red). Quantification of the percentage of p-PTK2 Y397 -positive cells in GFP-positive cells ( lower ). Arrowheads indicate the co-localization of the p-PTK2 Y397 with TDP-43-GFP-positive cells. Data are presented as the means ± SD of 3 independent experiments. **p<0.005 (Student’s t -test). Scale bars, 200 μm. (B) Immunocytochemistry was subsequently performed to detect ubiquitin (red) or DAPI (nuclei; blue). Quantification of the percentage of poly-ubiquitin-positive staining in GFP-positive cells ( lower ). Arrowheads indicate the co-localization of the poly-ubiquitin with TDP-43-GFP-positive cells. Data are presented as the means ± SD of 3 independent experiments. **p<0.005, ***p<0.001 (two-way ANOVA with Bonferroni multiple comparison test). Scale bars, 20 μm. (C) Immunocytochemistry was subsequently performed to detect the expression of cleaved caspase-3 (red) or DAPI (nuclei; blue). Quantification of the percentage of cleaved caspase-3-positive staining in GFP-positive cells (lower). Arrowheads indicate the co-localization of the caspase-3 with TDP-43-GFP-positive cells. Data are presented as the means ± SD of 3 independent experiments. **p<0.005, ***p<0.001 (two-way ANOVA with Bonferroni multiple comparison test). Scale bars, 200 μm.

Article Snippet: The following antibodies were used for immunocytochemistry: rabbit anti-cleaved caspase-3 (catalog no. 9611) [Cell Signaling Technology]; mouse anti-poly-ubiquitin (FK1) (catalog no. BML-PW8805) [Enzo Life Science]; rabbit anti-phospho-PTK2 (Tyr397) (catalog no. ab81298) [Abcam]; mouse anti-DDK (catalog no. TA50011) [Origene]; and rabbit anti-phospho-p62 (Ser403) (catalog no. GTX128171) [GeneTex].

Techniques: Transfection, Plasmid Preparation, Expressing, Construct, Immunocytochemistry, Staining

Journal: bioRxiv

Article Title: PTK2 regulates the UPS impairment via p62 phosphorylation in TDP-43 proteinopathy

doi: 10.1101/355446

Figure Lengend Snippet: A-B , The p62-, and p62 S403A -overexpressing N2a stable cells lines treated with MG132 (5 μm) for 24 h. Immunocytochemistry was performed thereafter. (A) Immunocytochemistry to detect the Cyto-ID autophagy dye (green) or ubiquitin (red) was subsequently performed. Quantification of the percentage of poly-ubiquitin-positive staining in all cells ( left ), the percentage of CytoID-positive and poly-ubiquitin-positive staining in poly-ubiquitin-positive cells ( middle ), or the percentage of CytoID-positive staining in total cells ( right ). Arrowheads indicate the colocalization of the Cyto-ID autophagy dye with poly-ubiquitin in the p62 stable cells lines. Data are presented as the means ± SD of 3 independent experiments. ***p<0.001, n.s. not significant (Student’s t -test). Scale bars, 10 μm. (B) Immunocytochemistry to detect the DDK (p62; green) or poly-ubiquitin (red) was subsequently performed. Quantification of the percentage of p62-positive puncta in poly-ubiquitin-positive cells (lower). Arrowheads indicate the co-localization of p62 with poly-ubiquitin in the p62 stable cells lines. Data are presented as the means ± SD of 3 independent experiments. *p<0.05 (Student’s t -test). Scale bars, 10 μm. C-D , p62-or p62 S403A -overexpressing N2a stable cells lines were transiently transfected with the control vector (pCMV6-AC-GFP) or TDP-43-expression construct (pCMV6-AC-TDP-43-GFP) for 3 days. Immunostaining was performed thereafter. (C) Immunocytochemistry was subsequently performed to detect poly-ubiquitin (red) or DAPI (nuclei; blue). Arrowheads indicate poly-ubiquitin-positive inclusions. Quantification of the percentage of poly-ubiquitin-positive staining in GFP-positive cells ( right ). Data are presented as the means ± SD of 3 independent experiments. **p<0.005 (two-way ANOVA with Bonferroni multiple comparison test). Scale bars, 10 μm. (D) Immunocytochemistry to detect caspase-3 (red) or DAPI (nuclei; blue) was subsequently performed. Arrowheads indicate caspase-3-positive cells. Quantification of the percentage of caspase-3-positive staining in GFP-positive cells (right). Data are presented as the means ± SD of 3 independent experiments. **p<0.005 (two-way ANOVA with Bonferroni multiple comparison test). Scale bars, 10 μm.

Article Snippet: The following antibodies were used for immunocytochemistry: rabbit anti-cleaved caspase-3 (catalog no. 9611) [Cell Signaling Technology]; mouse anti-poly-ubiquitin (FK1) (catalog no. BML-PW8805) [Enzo Life Science]; rabbit anti-phospho-PTK2 (Tyr397) (catalog no. ab81298) [Abcam]; mouse anti-DDK (catalog no. TA50011) [Origene]; and rabbit anti-phospho-p62 (Ser403) (catalog no. GTX128171) [GeneTex].

Techniques: Immunocytochemistry, Staining, Transfection, Plasmid Preparation, Expressing, Construct, Immunostaining