- PA5-101832 - Invitrogen Antibodies PTPRZ1 antibody

Su H, Fan S, Zhang L, Qi H
Frontiers in cellular neuroscience 2021;15:569424

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of PTPRZ in A549 cells. Samples were incubated with PTPRZ polyclonal antibody (Product # PA5-101832).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of PTPRZ in HUVEC cell lysate (Lane 1: treated with blocking peptide). Samples were incubated with PTPRZ polyclonal antibody (Product # PA5-101832).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of PTPRZ in A549 cells. Samples were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100, blocked with 10% serum (45 min at 25°C) incubated with PTPRZ polyclonal antibody (Product # PA5-101832) using a dilution of 1:200 (1 hr, 37°C), and followed by goat anti-rabbit IgG Alexa Fluor 594 at a dilution of 1:600.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of PTPRZ in A549 cells. Samples were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100, blocked with 10% serum (45 min at 25°C) incubated with PTPRZ polyclonal antibody (Product # PA5-101832) using a dilution of 1:200 (1 hr, 37°C), and followed by goat anti-rabbit IgG Alexa Fluor 594 at a dilution of 1:600.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Proliferation of reactive astrocytes and glial scarring is promoted by TMAO. (A) Evaluation of neurological function on MCAO/R rats treated with TMAO. (B) Number of collected chow pellets detected by staircase test in MCAO/R rats treated with TMAO. (C) Scoring results of asymmetry by cylinder test in MCAO/R rats treated with TMAO. (D) Statistical results of TMAO concentration in brain tissues of MCAO/R rats treated with TMAO. (E) Expression of GFAP, Neurocan, and Phophacan detected by immunofluorescence assay in brain tissues of MCAO/R rats treated with TMAO. (F) Expression of GFAP, Neurocan, and Phosphacan determined by Western blotting in brain tissues of MCAO/R rats treated with TMAO. * p < 0.05 compared with sham-operated rats ( n = 15). # p < 0.05 compared with MCAO/R rats ( n = 15). NS p > 0.05 compared with the control rats ( n = 15). One-way ANOVA is employed for data comparison among multiple groups followed by Tukey''s post hoc test.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 Overexpression of Smurf2 suppresses the promoting effect of TMAO on proliferation, migration, and viability of astrocytes. (A) Smurf2 expression in rat brain tissues ( n = 15) detected by IHC assay. (B) Smurf2 expression in differently treated cells determined by RT-qPCR. (C) Smurf2 expression determined by western blotting in differently treated cells. (D) Results of CCK-8 assay on cell viability upon different treatments. (E) Proliferation of cells upon different treatments detected by EdU assay. (F) Migration ability of cells upon different treatments assessed by Transwell migration assay. (G) Protein expression of GFAP, Neurocan and Phosphacan determined using Western blotting in differently treated cells. * p < 0.05 compared with the control group. # p < 0.05 compared with the TMAO + oe-NC group. One-way ANOVA was employed for data comparison among multiple groups followed by Tukey''s post hoc test.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Overexpression of ALK5 disrupts Smurf2-induced inhibition of astrocyte proliferation, migration and viability. (A) Expression of Smurf2 and ALK5 determined by Western blotting in each group of cells. (B) CCK-8 assay results of cell viability in each group. (C) EdU assay results of cell proliferation ability of each group. (D) Migration ability measured by Transwell migration assay in cells. (E) The expression of GFAP, Neurocan and Phosphacan in each group of cells detected by Western blotting. * p < 0.05 compared with the TMAO + oe-NC group. # p < 0.05 compared with the TMAO + oe-Smurf2 group. NS, p < 0.05 compared with the TMAO group. One-way ANOVA is employed for data comparison among multiple groups followed by Tukey''s post hoc test.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Silencing of ALK5 attenuates the promoting effect of TMAO on astrocyte proliferation, migration and viability. (A) Transfection efficiency of ALK5 in cells examined using RT-qPCR. (B) Protein expression of ALK5 in cells analyzed using Western blotting. (C) Expression of ALK5 and Smurf2 in cells determined by Western blotting. (D) Cell viability evaluated using CCK-8 assay. (E) Cell proliferation evaluated using EdU assay. (F) , Cell migration evaluated using Transwell assay. (G) , The expression of GFAP, Neurocan and Phosphacan in each group of cells detected by Western blotting. * p < 0.05 compared with the sh-NC group or the TMAO + oe-NC group. NS, p > 0.05 compared with the TMAO group. Unpaired t -test is adopted for data comparison between two groups. One-way ANOVA is employed for data comparison among multiple groups followed by Tukey''s post hoc test.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 TMAO promotes reactive astrocyte proliferation and glial scarring through the Smurf2/ALK5 axis. (A) Smurf2 and ALK5 expression determined by western blotting in brain tissues of MCAO/R rats. (B) Neurological scoring of rats after different treatments. (C) The number of collected chow pellets by rats in each group detected by staircase test. (D) Asymmetric function detected by cylinder test. (E) Expression of GFAP, Neurocan and Phosphacan detected by immunofluorescence assay in brain tissues of rats after different treatments. (F) Protein expression of GFAP, Neurocan and Phosphacan determined using Western blotting in brain tissues of rats after different treatments. * p < 0.05 compared with MCAO/R + TMAO + sh-NC group ( n = 15). NS p > 0.05 compared with MCAO/R + TMAO group ( n = 15). One-way ANOVA is employed for data comparison among multiple groups followed by Tukey''s post hoc test.

PA5-101832