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

Journal: Neuron

Article Title: Targeted proteoform mapping uncovers specific Neurexin-3 variants required for dendritic inhibition

doi: 10.1016/j.neuron.2022.04.017

Figure Lengend Snippet:

Article Snippet: The following antibodies were used in this study: rabbit polyclonal anti-β-actin (Abcam; Cat# ab8227; RRID: AB_2305186; LOT# GR3314266-1), mouse monoclonal anti-calbindin (Swant; Cat# 300; RRID: AB_10000347; LOT# 17 (F)), goat polyclonal anti-calretinin (Swant; Cat# CG1; RRID: AB_10000342; LOT# 1§.1), mouse monoclonal anti-CamKII alpha (Thermo Fisher Scientific; 6G9; Cat# Ma1-048; RRID: AB_325403; LOT# TH269517), mouse monoclonal anti-cannabinoid receptor 1 (Immunogene; IMG-3C2; Cat# IMG-CB1R-mAb001; LOT# CJ03), guinea pig polyclonal anti-cholecystokinin (Synaptic Systems, Cat# 438004; RRID: AB_2814938; LOT# 1-1), mouse monoclonal anti-GAD67 (Millipore; 1G10.2; Cat# MAB5406; RRID: AB_2278725; LOT# 3015328), rabbit polyclonal anti-GAPDH (Enogene; Cat# E1C604; LOT# R14Q12), mouse monoclonal anti-gephyrin (Synaptic Systems; mAb7a; Cat# 147021; RRID: AB_2232546; LOT# 147021/15), mouse monoclonal anti-gephyrin (Synaptic Systems; mAb7a; Cat# 147011; RRID: AB_887717; LOT# 147011/54), rat monoclonal anti-HA (Roche; 3F10; Cat# 11867431001; RRID: AB_390919; LOT# 34502100), rabbit monoclonal anti-HA (Cell Signaling; 3724; Cat# 3724; RRID: AB_1549585; LOT# 9), guinea pig polyclonal anti-MAP2 (Synaptic Systems; Cat# 188004; RRID: AB_2138181; LOT# 2-26), mouse monoclonal anti-MAP2 (Synaptic Systems; 198A5; Cat# 188011; RRID: AB_2147096; LOT# 1-10), rabbit polyclonal anti-neurexin , chicken anti-neurexin , rabbit anti-neuroligin , rabbit monoclonal anti-nNOS (Cell Signaling; C7D7; Cat# 4231; RRID: AB_2152485; LOT# 2), goat polyclonal anti-parvalbumin (Swant, Cat# PVG214; RRID: AB_10000345), mouse monoclonal anti-PSD95 (Santa Cruz; 7E3; Cat# sc32290; RRID: AB_628114; LOT# J1509), goat polyclonal anti-somatostatin (Santa Cruz; Cat# sc7819; RRID: AB_2302603; LOT# L1611), mouse monoclonal anti-Synaptotagmin 2 (Zebrafish International Resource Center; Cat# znp-1; RRID: AB_10013783), mouse monoclonal anti-V5 (Biorad; SV5-PK1; Cat# MCA1360; RRID: AB_322378; LOT# 148239), guinea pig polyclonal anti-vGAT (Synaptic Systems; Cat# 131004; RRID: AB_887873; LOT# 2-42), guinea pig polyclonal anti-vGlut1 (Millipore; Cat# AB5905; RRID: AB_2301751; LOT# 3308226), rabbit polyclonal anti-VIP (Immunostar; Cat# 20077; RRID: AB_572270; LOT# 1513001), guinea pig anti-somatostatin (raised in the present study against amino acid residues 35-88 of mouse pro-somatostatin, GenBank: #BC010770.1), goat anti-cannabinoid receptor 1 (Nittobo Medical, MSFR100600; RRID: AB_2571592), guinea pig anti-GABAA receptor alpha 1 (Nittobo Medical, MSFR101540; RRID: AB_2571572), goat anti-vGAT (Nittobo Medical, MSFR106130; RRID: AB_2571623), guinea pig anti-PSD95 (Nittobo Medical, MSFR105180; RRID: AB_2571612).

Techniques: Recombinant, Reporter Assay, Multiplex Assay, Plasmid Preparation, Software, Microscopy