bs 1369r  (Bioss)

Journal: Neural Regeneration Research

Article Title: Hypoxia-preconditioned bone marrow–derived mesenchymal stem cells protect neurons from cardiac arrest–induced pyroptosis

doi: 10.4103/NRR.NRR-D-23-01922

Figure Lengend Snippet: HP-BMSCs suppress OGD-induced neuronal pyroptosis to some extent by inhibiting the MAPK and NF-κB pathways. (A) From left to right: BMSCs at passage 3 (P3), osteogenic differentiation BMSCs, and adipogenic differentiation BMSCs. The cells are typically spindle-shaped. Scale bars: 100 μm. (B) Flow cytometric analysis showing the expression of BMSC surface markers. The cells expressed CD29 and CD90 but did not express CD11b and CD45. (C) Immunofluorescence staining of primary cultured neurons for MAP2 (red, Alexa Fluor 594), whereas nuclei were labeled with DAPI (blue). Scale bars: 75 μm. (D) Western blotting analysis was performed to evaluate the expression of NLRP3, GSDMD-N, ASC, cleaved-caspase-1, NF-κB, JNK, and p38 MAPK in primary neurons from the indicated group ( n = 3). The results of parallel test were shown in Additional Figure 2 . (E–K) Quantitative analysis of NLRP3 (E), GSDMD-N (F), ASC (G), cleaved-caspase-1 (H), NF-κB (I), JNK (J), and p38 MAPK (K) protein expression levels as shown in D. Results are expressed as mean ± SD. * P < 0.05, vs . control group; # P < 0.05, vs . OGD group; & P < 0.05, vs . OGD + N-BMSCs group; $ P < 0.05, vs . OGD + HP-BMSCs group (one-way analysis of variance followed by Tukey’s multiple comparisons test). BMSCs: Bone marrow mesenchymal stem cells; HP-BMSCs: hypoxia preconditioned bone marrow mesenchymal stem cells; N-BMSCs: normal-cultured BMSCs; OGD: oxygen-glucose deprivation.

Article Snippet: Finally, mature neurons were identified by staining with the neuronal marker microtubule-associated protein 2 (MAP2, rabbit, 1:200, Bioss, Cat# bs-1369R; RRID: AB_10853045).

Techniques: Expressing, Immunofluorescence, Staining, Cell Culture, Labeling, Western Blot, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Reducing polypyrimidine tract‑binding protein 1 fails to promote neuronal transdifferentiation on HT22 and mouse astrocyte cells under physiological conditions

doi: 10.3892/etm.2023.12360

Figure Lengend Snippet: Effect of a 3-day decrease in PTBP1 protein level in HT22 cells on cell differentiation and maturation. (A) Representative western blotting of MAP2, PTBP1, βIII-Tubulin, NeuN and GAPDH (n=4 per group). Quantitative statistical analysis results of (B) MAP2, (C) PTBP1, (D) βIII-Tubulin, (E) NeuN. (F) Representative immunofluorescence staining images and quantitative statistical analysis results of (G) βIII-Tubulin and (H) PTBP1. (I) Representative immunofluorescence staining images and quantitative statistical analysis results of (J) NeuN and (K) PTBP1. (L) Representative immunofluorescence staining images and quantitative statistical analysis results of (M) MAP2 and (N) PTBP1 (n=3 per group). Scar bar, 50 µm. ** P<0.01, *** P<0.001 and **** P<0.0001. PTBP1, polypyrimidine tract-binding protein 1; MAP2, microtubule-associated protein 2; NC, negative control; si, small interfering; ns, not significant.

Article Snippet: After blocking for 1 h with 3% bovine serum albumin (cat. no. GC305010; Wuhan Servicebio Technology Co., Ltd.) at room temperature, the membrane was incubated overnight at 4˚C with appropriate primary antibodies: Rabbit anti-MAP2 antibody (1:1,000; cat. no. bs-1369R; BIOSS), rabbit anti-PTBP1 antibody (1:2,000; cat. no. 101043-T46; Sino Biological, Inc.), mouse anti-βIII-Tubulin antibody (1:1,000; cat. no. Sc-80016; Santa Cruz Biotechnology, Inc.), rabbit anti-NeuN antibody (1:1,000; cat. no. AF1072, Beyotime Institute of Biotechnology) and mouse anti-GAPDH antibody (cat. no. GB15002, Wuhan Servicebio Technology Co., Ltd.).

Techniques: Cell Differentiation, Western Blot, Immunofluorescence, Staining, Binding Assay, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: Reducing polypyrimidine tract‑binding protein 1 fails to promote neuronal transdifferentiation on HT22 and mouse astrocyte cells under physiological conditions

doi: 10.3892/etm.2023.12360

Figure Lengend Snippet: Effect of a 5 day decrease in PTBP1 protein level in HT22 cells on cell differentiation and maturation. (A) Representative western blotting of MAP2, PTBP1, βIII-Tubulin, NeuN and GAPDH (n=4 per group). Quantitative statistical analysis results of (B) MAP2, (C) PTBP1, (D) βIII-Tubulin and (E) NeuN. (F) Representative immunofluorescence staining images and quantitative statistical analysis results of (G) βIII-Tubulin and (H) PTBP1. (I) Representative immunofluorescence staining images and quantitative statistical analysis results of (J) NeuN and (K) PTBP1. (L) Representative immunofluorescence staining images and quantitative statistical analysis results of (M) MAP2 and (N) PTBP1 (n=3 per group). Scar bar, 50 µm. ** P<0.01, *** P<0.001 and **** P<0.0001. PTBP1, polypyrimidine tract-binding protein 1; MAP2, microtubule-associated protein 2; NC, negative control; si, small interfering; ns, not significant.

Article Snippet: After blocking for 1 h with 3% bovine serum albumin (cat. no. GC305010; Wuhan Servicebio Technology Co., Ltd.) at room temperature, the membrane was incubated overnight at 4˚C with appropriate primary antibodies: Rabbit anti-MAP2 antibody (1:1,000; cat. no. bs-1369R; BIOSS), rabbit anti-PTBP1 antibody (1:2,000; cat. no. 101043-T46; Sino Biological, Inc.), mouse anti-βIII-Tubulin antibody (1:1,000; cat. no. Sc-80016; Santa Cruz Biotechnology, Inc.), rabbit anti-NeuN antibody (1:1,000; cat. no. AF1072, Beyotime Institute of Biotechnology) and mouse anti-GAPDH antibody (cat. no. GB15002, Wuhan Servicebio Technology Co., Ltd.).

Techniques: Cell Differentiation, Western Blot, Immunofluorescence, Staining, Binding Assay, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: Reducing polypyrimidine tract‑binding protein 1 fails to promote neuronal transdifferentiation on HT22 and mouse astrocyte cells under physiological conditions

doi: 10.3892/etm.2023.12360

Figure Lengend Snippet: Effect of a 3 day decrease in PTBP1 protein level in MA cells on cell differentiation. (A) Representative western blotting of MAP2, PTBP1, βIII-Tubulin, NeuN and GAPDH (n=4 per group). Quantitative statistical analysis results of (B) MAP2, (C) PTBP1, (D) βIII-Tubulin and (E) NeuN. (F) Representative immunofluorescence staining images and quantitative statistical analysis results of (G) βIII-Tubulin and (H) PTBP1. (I) Representative immunofluorescence staining images and quantitative statistical analysis results of (J) NeuN and (K) PTBP1. (L) Representative immunofluorescence staining images and quantitative statistical analysis results of (M) MAP2 and (N) PTBP1 (n=3 per group). Scar bar, 50 µm. *** P<0.001 and **** P<0.0001. PTBP1, polypyrimidine tract-binding protein 1; MAP2, microtubule-associated protein 2; NC, negative control; si, small interfering; ns, not significant; MA, mouse astrocyte.

Article Snippet: After blocking for 1 h with 3% bovine serum albumin (cat. no. GC305010; Wuhan Servicebio Technology Co., Ltd.) at room temperature, the membrane was incubated overnight at 4˚C with appropriate primary antibodies: Rabbit anti-MAP2 antibody (1:1,000; cat. no. bs-1369R; BIOSS), rabbit anti-PTBP1 antibody (1:2,000; cat. no. 101043-T46; Sino Biological, Inc.), mouse anti-βIII-Tubulin antibody (1:1,000; cat. no. Sc-80016; Santa Cruz Biotechnology, Inc.), rabbit anti-NeuN antibody (1:1,000; cat. no. AF1072, Beyotime Institute of Biotechnology) and mouse anti-GAPDH antibody (cat. no. GB15002, Wuhan Servicebio Technology Co., Ltd.).

Techniques: Cell Differentiation, Western Blot, Immunofluorescence, Staining, Binding Assay, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: Reducing polypyrimidine tract‑binding protein 1 fails to promote neuronal transdifferentiation on HT22 and mouse astrocyte cells under physiological conditions

doi: 10.3892/etm.2023.12360

Figure Lengend Snippet: Effect of a 5 day decrease in PTBP1 protein level in MA cells on cell differentiation. (A) Representative western blotting of MAP2, PTBP1, βIII-Tubulin, NeuN and GAPDH (n=4 per group). Quantitative statistical analysis results of (B) MAP2, (C) PTBP1, (D) βIII-Tubulin and (E) NeuN. (F) Representative immunofluorescence staining images and quantitative statistical analysis results of (G) βIII-Tubulin and (H) PTBP1. (I) Representative immunofluorescence staining images and quantitative statistical analysis results of (J) NeuN and (K) PTBP1. (L) Representative immunofluorescence staining images and quantitative statistical analysis results of (M) MAP2 and (N) PTBP1 (n=3 per group). Scar bar, 50 µm. *** P<0.001 and **** P<0.0001. PTBP1, polypyrimidine tract-binding protein 1; MAP2, microtubule-associated protein 2; NC, negative control; si, small interfering; ns, not significant; MA, mouse astrocyte.

Article Snippet: After blocking for 1 h with 3% bovine serum albumin (cat. no. GC305010; Wuhan Servicebio Technology Co., Ltd.) at room temperature, the membrane was incubated overnight at 4˚C with appropriate primary antibodies: Rabbit anti-MAP2 antibody (1:1,000; cat. no. bs-1369R; BIOSS), rabbit anti-PTBP1 antibody (1:2,000; cat. no. 101043-T46; Sino Biological, Inc.), mouse anti-βIII-Tubulin antibody (1:1,000; cat. no. Sc-80016; Santa Cruz Biotechnology, Inc.), rabbit anti-NeuN antibody (1:1,000; cat. no. AF1072, Beyotime Institute of Biotechnology) and mouse anti-GAPDH antibody (cat. no. GB15002, Wuhan Servicebio Technology Co., Ltd.).

Techniques: Cell Differentiation, Western Blot, Immunofluorescence, Staining, Binding Assay, Negative Control