wt rats Search Results


rat anti mouse cd18  (Cedarlane)
85
Cedarlane rat anti mouse cd18
Rat Anti Mouse Cd18, supplied by Cedarlane, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rat anti mouse cd18/product/Cedarlane
Average 85 stars, based on 1 article reviews
rat anti mouse cd18 - by Bioz Stars, 2026-05
85/100 stars
  Buy from Supplier
cd18  (Cedarlane)
93
Cedarlane cd18
Cd18, supplied by Cedarlane, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/cd18/product/Cedarlane
Average 93 stars, based on 1 article reviews
cd18 - by Bioz Stars, 2026-05
93/100 stars
  Buy from Supplier
erk2  (Addgene inc)
92
Addgene inc erk2
Figure 1. Overexpression of ERK1/2 downregulates GABAB receptors. HEK 293 cells were trans- fected with GABAB1, GABAB2 and empty vector or EGFP as a control (Ctrl) or with GABAB1, GABAB2 and either ERK1 or <t>ERK2.</t> After 2 days, the cells were tested for ERK1/2 and GABAB receptor ex- pression by immunofluorescence staining using antibodies directed against ERK1/2 and GABAB1 or GABAB2. (A) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB1. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 101–148 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001). (B) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB2. Left: rep- resentative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 54–95 cells per condition from four independent experiments. One Way ANOVA with Tukey’s post-test (ns, p > 0.05; ****, p < 0.0001). (C) Transfection with ERK1 or ERK2 downregulated the cell surface expression of GABAB receptors as probed with antibodies directed against an extracellular located epitope in the N-terminal domain of GABAB2. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 84–113 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001).
Erk2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/erk2/product/Addgene inc
Average 92 stars, based on 1 article reviews
erk2 - by Bioz Stars, 2026-05
92/100 stars
  Buy from Supplier
wt rats  (SynGap Research Fund Inc)
90
SynGap Research Fund Inc wt rats
Figure 1. Overexpression of ERK1/2 downregulates GABAB receptors. HEK 293 cells were trans- fected with GABAB1, GABAB2 and empty vector or EGFP as a control (Ctrl) or with GABAB1, GABAB2 and either ERK1 or <t>ERK2.</t> After 2 days, the cells were tested for ERK1/2 and GABAB receptor ex- pression by immunofluorescence staining using antibodies directed against ERK1/2 and GABAB1 or GABAB2. (A) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB1. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 101–148 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001). (B) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB2. Left: rep- resentative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 54–95 cells per condition from four independent experiments. One Way ANOVA with Tukey’s post-test (ns, p > 0.05; ****, p < 0.0001). (C) Transfection with ERK1 or ERK2 downregulated the cell surface expression of GABAB receptors as probed with antibodies directed against an extracellular located epitope in the N-terminal domain of GABAB2. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 84–113 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001).
Wt Rats, supplied by SynGap Research Fund Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/wt rats/product/SynGap Research Fund Inc
Average 90 stars, based on 1 article reviews
wt rats - by Bioz Stars, 2026-05
90/100 stars
  Buy from Supplier
male wt and ws/ws rats  (Japan SLC inc)
90
Japan SLC inc male wt and ws/ws rats
Figure 1. Overexpression of ERK1/2 downregulates GABAB receptors. HEK 293 cells were trans- fected with GABAB1, GABAB2 and empty vector or EGFP as a control (Ctrl) or with GABAB1, GABAB2 and either ERK1 or <t>ERK2.</t> After 2 days, the cells were tested for ERK1/2 and GABAB receptor ex- pression by immunofluorescence staining using antibodies directed against ERK1/2 and GABAB1 or GABAB2. (A) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB1. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 101–148 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001). (B) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB2. Left: rep- resentative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 54–95 cells per condition from four independent experiments. One Way ANOVA with Tukey’s post-test (ns, p > 0.05; ****, p < 0.0001). (C) Transfection with ERK1 or ERK2 downregulated the cell surface expression of GABAB receptors as probed with antibodies directed against an extracellular located epitope in the N-terminal domain of GABAB2. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 84–113 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001).
Male Wt And Ws/Ws Rats, supplied by Japan SLC inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/male wt and ws/ws rats/product/Japan SLC inc
Average 90 stars, based on 1 article reviews
male wt and ws/ws rats - by Bioz Stars, 2026-05
90/100 stars
  Buy from Supplier
male wt and ws/ws rats  (Hamamatsu)
90
Hamamatsu male wt and ws/ws rats
Figure 1. Overexpression of ERK1/2 downregulates GABAB receptors. HEK 293 cells were trans- fected with GABAB1, GABAB2 and empty vector or EGFP as a control (Ctrl) or with GABAB1, GABAB2 and either ERK1 or <t>ERK2.</t> After 2 days, the cells were tested for ERK1/2 and GABAB receptor ex- pression by immunofluorescence staining using antibodies directed against ERK1/2 and GABAB1 or GABAB2. (A) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB1. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 101–148 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001). (B) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB2. Left: rep- resentative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 54–95 cells per condition from four independent experiments. One Way ANOVA with Tukey’s post-test (ns, p > 0.05; ****, p < 0.0001). (C) Transfection with ERK1 or ERK2 downregulated the cell surface expression of GABAB receptors as probed with antibodies directed against an extracellular located epitope in the N-terminal domain of GABAB2. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 84–113 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001).
Male Wt And Ws/Ws Rats, supplied by Hamamatsu, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/male wt and ws/ws rats/product/Hamamatsu
Average 90 stars, based on 1 article reviews
male wt and ws/ws rats - by Bioz Stars, 2026-05
90/100 stars
  Buy from Supplier
wt sd rats  (Cyagen Biosciences)
90
Cyagen Biosciences wt sd rats
Figure 1. Overexpression of ERK1/2 downregulates GABAB receptors. HEK 293 cells were trans- fected with GABAB1, GABAB2 and empty vector or EGFP as a control (Ctrl) or with GABAB1, GABAB2 and either ERK1 or <t>ERK2.</t> After 2 days, the cells were tested for ERK1/2 and GABAB receptor ex- pression by immunofluorescence staining using antibodies directed against ERK1/2 and GABAB1 or GABAB2. (A) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB1. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 101–148 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001). (B) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB2. Left: rep- resentative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 54–95 cells per condition from four independent experiments. One Way ANOVA with Tukey’s post-test (ns, p > 0.05; ****, p < 0.0001). (C) Transfection with ERK1 or ERK2 downregulated the cell surface expression of GABAB receptors as probed with antibodies directed against an extracellular located epitope in the N-terminal domain of GABAB2. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 84–113 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001).
Wt Sd Rats, supplied by Cyagen Biosciences, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/wt sd rats/product/Cyagen Biosciences
Average 90 stars, based on 1 article reviews
wt sd rats - by Bioz Stars, 2026-05
90/100 stars
  Buy from Supplier
wt rats  (Harlan Laboratories)
90
Harlan Laboratories wt rats
Figure 1. Overexpression of ERK1/2 downregulates GABAB receptors. HEK 293 cells were trans- fected with GABAB1, GABAB2 and empty vector or EGFP as a control (Ctrl) or with GABAB1, GABAB2 and either ERK1 or <t>ERK2.</t> After 2 days, the cells were tested for ERK1/2 and GABAB receptor ex- pression by immunofluorescence staining using antibodies directed against ERK1/2 and GABAB1 or GABAB2. (A) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB1. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 101–148 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001). (B) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB2. Left: rep- resentative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 54–95 cells per condition from four independent experiments. One Way ANOVA with Tukey’s post-test (ns, p > 0.05; ****, p < 0.0001). (C) Transfection with ERK1 or ERK2 downregulated the cell surface expression of GABAB receptors as probed with antibodies directed against an extracellular located epitope in the N-terminal domain of GABAB2. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 84–113 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001).
Wt Rats, supplied by Harlan Laboratories, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/wt rats/product/Harlan Laboratories
Average 90 stars, based on 1 article reviews
wt rats - by Bioz Stars, 2026-05
90/100 stars
  Buy from Supplier
rip1 d138n and wt littermate rats  (Genentech inc)
90
Genentech inc rip1 d138n and wt littermate rats
Figure 1. Overexpression of ERK1/2 downregulates GABAB receptors. HEK 293 cells were trans- fected with GABAB1, GABAB2 and empty vector or EGFP as a control (Ctrl) or with GABAB1, GABAB2 and either ERK1 or <t>ERK2.</t> After 2 days, the cells were tested for ERK1/2 and GABAB receptor ex- pression by immunofluorescence staining using antibodies directed against ERK1/2 and GABAB1 or GABAB2. (A) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB1. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 101–148 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001). (B) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB2. Left: rep- resentative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 54–95 cells per condition from four independent experiments. One Way ANOVA with Tukey’s post-test (ns, p > 0.05; ****, p < 0.0001). (C) Transfection with ERK1 or ERK2 downregulated the cell surface expression of GABAB receptors as probed with antibodies directed against an extracellular located epitope in the N-terminal domain of GABAB2. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 84–113 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001).
Rip1 D138n And Wt Littermate Rats, supplied by Genentech inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rip1 d138n and wt littermate rats/product/Genentech inc
Average 90 stars, based on 1 article reviews
rip1 d138n and wt littermate rats - by Bioz Stars, 2026-05
90/100 stars
  Buy from Supplier
parturient sprague dawley (sd) rats (d000017, wt/wt)  (Pharmatech)
90
Pharmatech parturient sprague dawley (sd) rats (d000017, wt/wt)
Figure 1. Overexpression of ERK1/2 downregulates GABAB receptors. HEK 293 cells were trans- fected with GABAB1, GABAB2 and empty vector or EGFP as a control (Ctrl) or with GABAB1, GABAB2 and either ERK1 or <t>ERK2.</t> After 2 days, the cells were tested for ERK1/2 and GABAB receptor ex- pression by immunofluorescence staining using antibodies directed against ERK1/2 and GABAB1 or GABAB2. (A) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB1. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 101–148 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001). (B) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB2. Left: rep- resentative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 54–95 cells per condition from four independent experiments. One Way ANOVA with Tukey’s post-test (ns, p > 0.05; ****, p < 0.0001). (C) Transfection with ERK1 or ERK2 downregulated the cell surface expression of GABAB receptors as probed with antibodies directed against an extracellular located epitope in the N-terminal domain of GABAB2. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 84–113 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001).
Parturient Sprague Dawley (Sd) Rats (D000017, Wt/Wt), supplied by Pharmatech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/parturient sprague dawley (sd) rats (d000017, wt/wt)/product/Pharmatech
Average 90 stars, based on 1 article reviews
parturient sprague dawley (sd) rats (d000017, wt/wt) - by Bioz Stars, 2026-05
90/100 stars
  Buy from Supplier
ldlr -ko wt sprague-dawley rats  (Harlan Laboratories)
90
Harlan Laboratories ldlr -ko wt sprague-dawley rats
Figure 1. Overexpression of ERK1/2 downregulates GABAB receptors. HEK 293 cells were trans- fected with GABAB1, GABAB2 and empty vector or EGFP as a control (Ctrl) or with GABAB1, GABAB2 and either ERK1 or <t>ERK2.</t> After 2 days, the cells were tested for ERK1/2 and GABAB receptor ex- pression by immunofluorescence staining using antibodies directed against ERK1/2 and GABAB1 or GABAB2. (A) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB1. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 101–148 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001). (B) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB2. Left: rep- resentative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 54–95 cells per condition from four independent experiments. One Way ANOVA with Tukey’s post-test (ns, p > 0.05; ****, p < 0.0001). (C) Transfection with ERK1 or ERK2 downregulated the cell surface expression of GABAB receptors as probed with antibodies directed against an extracellular located epitope in the N-terminal domain of GABAB2. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 84–113 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001).
Ldlr Ko Wt Sprague Dawley Rats, supplied by Harlan Laboratories, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ldlr -ko wt sprague-dawley rats/product/Harlan Laboratories
Average 90 stars, based on 1 article reviews
ldlr -ko wt sprague-dawley rats - by Bioz Stars, 2026-05
90/100 stars
  Buy from Supplier
wild-type (wt) lewis (lew) rats  (Harlan Laboratories)
90
Harlan Laboratories wild-type (wt) lewis (lew) rats
Figure 1. Overexpression of ERK1/2 downregulates GABAB receptors. HEK 293 cells were trans- fected with GABAB1, GABAB2 and empty vector or EGFP as a control (Ctrl) or with GABAB1, GABAB2 and either ERK1 or <t>ERK2.</t> After 2 days, the cells were tested for ERK1/2 and GABAB receptor ex- pression by immunofluorescence staining using antibodies directed against ERK1/2 and GABAB1 or GABAB2. (A) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB1. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 101–148 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001). (B) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB2. Left: rep- resentative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 54–95 cells per condition from four independent experiments. One Way ANOVA with Tukey’s post-test (ns, p > 0.05; ****, p < 0.0001). (C) Transfection with ERK1 or ERK2 downregulated the cell surface expression of GABAB receptors as probed with antibodies directed against an extracellular located epitope in the N-terminal domain of GABAB2. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 84–113 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001).
Wild Type (Wt) Lewis (Lew) Rats, supplied by Harlan Laboratories, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/wild-type (wt) lewis (lew) rats/product/Harlan Laboratories
Average 90 stars, based on 1 article reviews
wild-type (wt) lewis (lew) rats - by Bioz Stars, 2026-05
90/100 stars
  Buy from Supplier

Image Search Results


Figure 1. Overexpression of ERK1/2 downregulates GABAB receptors. HEK 293 cells were trans- fected with GABAB1, GABAB2 and empty vector or EGFP as a control (Ctrl) or with GABAB1, GABAB2 and either ERK1 or ERK2. After 2 days, the cells were tested for ERK1/2 and GABAB receptor ex- pression by immunofluorescence staining using antibodies directed against ERK1/2 and GABAB1 or GABAB2. (A) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB1. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 101–148 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001). (B) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB2. Left: rep- resentative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 54–95 cells per condition from four independent experiments. One Way ANOVA with Tukey’s post-test (ns, p > 0.05; ****, p < 0.0001). (C) Transfection with ERK1 or ERK2 downregulated the cell surface expression of GABAB receptors as probed with antibodies directed against an extracellular located epitope in the N-terminal domain of GABAB2. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 84–113 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001).

Journal: International journal of molecular sciences

Article Title: ERK1/2-Dependent Phosphorylation of GABA B1 (S867/T872), Controlled by CaMKIIβ, Is Required for GABA B Receptor Degradation under Physiological and Pathological Conditions.

doi: 10.3390/ijms241713436

Figure Lengend Snippet: Figure 1. Overexpression of ERK1/2 downregulates GABAB receptors. HEK 293 cells were trans- fected with GABAB1, GABAB2 and empty vector or EGFP as a control (Ctrl) or with GABAB1, GABAB2 and either ERK1 or ERK2. After 2 days, the cells were tested for ERK1/2 and GABAB receptor ex- pression by immunofluorescence staining using antibodies directed against ERK1/2 and GABAB1 or GABAB2. (A) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB1. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 101–148 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001). (B) Transfection with ERK1 or ERK2 downregulated the expression of total GABAB2. Left: rep- resentative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 54–95 cells per condition from four independent experiments. One Way ANOVA with Tukey’s post-test (ns, p > 0.05; ****, p < 0.0001). (C) Transfection with ERK1 or ERK2 downregulated the cell surface expression of GABAB receptors as probed with antibodies directed against an extracellular located epitope in the N-terminal domain of GABAB2. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescence intensities. The data represent the mean ± SD of 84–113 cells per condition from four independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001).

Article Snippet: The following plasmids were used for this study: HA-tagged GABAB1a [39]; GABAB2 [3]; HA-tagged GABAB1a(S867A) and HA-tagged GABAB1a(T872A) (mutations were custommade by GenScript, Piscataway NJ, USA), ERK1 (Addgene plasmid 12656, Watertown, MA, USA) [40], ERK2 (Addgene plasmid 40812) [40] and GFP-tagged CaMKIIβ (Addgene plasmid 21227) [41].

Techniques: Over Expression, Plasmid Preparation, Control, Staining, Transfection, Expressing

Figure 3. ERK1/2 and CaMKIIβ mediated phosphorylation of GABAB1 at serine 867 (S867) and threonine 872 (T872). (A) Transfection of neurons with mutant GABAB1 containing inactivated phosphorylation sites T872A or S867A upregulated the cell surface expression of GABAB receptors as determined by immunofluorescence staining using antibodies directed against the HA-tagged GABAB1 phospho-mutants. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescent intensities. The data represent the mean ± SD of 24 cells per condition from three independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001). (B) ERK1/2 and CaMKII mediated phosphorylation of S867 in GABAB1. HEK-293 cells were transfected with wild-type GABAB1/GABAB2 (wt) or with the phospho-mutant GABAB1(S867A)/GABAB2 with or without CaMKIIβ, ERK1 or ERK2 and tested for GABAB receptor phosphorylation by in situ PLA using antibodies directed against phospho-serine and HA-tagged

Journal: International journal of molecular sciences

Article Title: ERK1/2-Dependent Phosphorylation of GABA B1 (S867/T872), Controlled by CaMKIIβ, Is Required for GABA B Receptor Degradation under Physiological and Pathological Conditions.

doi: 10.3390/ijms241713436

Figure Lengend Snippet: Figure 3. ERK1/2 and CaMKIIβ mediated phosphorylation of GABAB1 at serine 867 (S867) and threonine 872 (T872). (A) Transfection of neurons with mutant GABAB1 containing inactivated phosphorylation sites T872A or S867A upregulated the cell surface expression of GABAB receptors as determined by immunofluorescence staining using antibodies directed against the HA-tagged GABAB1 phospho-mutants. Left: representative images (scale bar: 10 µm). Right: quantification of fluorescent intensities. The data represent the mean ± SD of 24 cells per condition from three independent experiments. Brown–Forsythe and Welch’s ANOVA with Games-Howell’s post-test (ns, p > 0.05; ****, p < 0.0001). (B) ERK1/2 and CaMKII mediated phosphorylation of S867 in GABAB1. HEK-293 cells were transfected with wild-type GABAB1/GABAB2 (wt) or with the phospho-mutant GABAB1(S867A)/GABAB2 with or without CaMKIIβ, ERK1 or ERK2 and tested for GABAB receptor phosphorylation by in situ PLA using antibodies directed against phospho-serine and HA-tagged

Article Snippet: The following plasmids were used for this study: HA-tagged GABAB1a [39]; GABAB2 [3]; HA-tagged GABAB1a(S867A) and HA-tagged GABAB1a(T872A) (mutations were custommade by GenScript, Piscataway NJ, USA), ERK1 (Addgene plasmid 12656, Watertown, MA, USA) [40], ERK2 (Addgene plasmid 40812) [40] and GFP-tagged CaMKIIβ (Addgene plasmid 21227) [41].

Techniques: Phospho-proteomics, Transfection, Mutagenesis, Expressing, Staining, In Situ