In situ hybridization of Grik1 and Grm6 and immunostaining of GRM6 upon loss of Fat3 in mouse retina (Related toFig. 7). (A–A″) VSX2 immunostaining (A) after in situ hybridization for Grik1 (A′) and Grm6 (A″) in WT retina. (B–B″) VSX2 immunostaining (B) after in situ hybridization for Grik1 (B′) and Grm6 (B″) in Fat3∆TM/∆TM retina. (C) Quantification of Grik1 RNA mean fluorescence intensity in bipolar cells. WT: 1.00 ± 0.03, n = 4 animals, 16 retinal regions; Fat3∆TM/∆TM: 0.75 ± 0.04, n = 4 animals, 17 retinal regions. Each data point corresponds to a retinal region, color-coded by animal, analyzed using a nested two-tailed test. (D) Quantification of Grm6 RNA mean fluorescence intensity in bipolar cells. WT: 1.00 ± 0.02, n = 4 animals, from 16 retinal regions; Fat3∆TM/∆TM: 0.98 ± 0.02, n = 4 animals, from 17 retinal regions. Each data point corresponds to a retinal region, color-coded by animal, analyzed using a nested two-tailed test. (E) Immunostaining for GRM6 in WT retinas. (F) Immunostaining for GRM6 in Fat3∆TM/∆TM retinas. (E′ and F′) Cone arrestin (ARR3) labels the cone photoreceptors endings in the OPL in E′ and F′. (G) Immunostaining for GRM6 in WT retinas, littermates of Fat3∆ICD-GFP/∆ICD-GFP animals. (H) Immunostaining for GRM6 in Fat3∆ICD-GFP/∆ICD-GFP retinas. Cone arrestin (ARR3) labels the cone photoreceptors endings in the OPL in G′ and H′. (I) Quantification of GRM6 integrated intensity in the OPL. WT controls: 1.00 ± 0.08 (n = 8 animals, 35 retinal regions); Fat3∆TM/∆TM: 1.15 ± 0.09 (n = 9 animals, 41 retinal regions). Each data point corresponds to a retinal region, color-coded by animal, nested two-tailed test. (J) Quantification of GRM6 integrated intensity in the OPL. WT Controls: 1.00 ± 0.09 (n = 5 animals, 21 retinal regions); Fat3∆ICD/∆ICD: 0.86 ± 0.07 (n = 5 animals, 22 retinal regions). Each data point corresponds to a retinal region, color-coded by animal, analyzed using a nested two-tailed test. (K) Flicker ERG amplitude at 30 Hz for WT (18.70 ± 2.05 µV n = 6 eyes) and Ptprs−/− retinas (24.50 ± 2.86 µV, n = 6 eyes). (L) Flicker ERG implicit time at 20 Hz for WT (34.3 ± 0.6 ms, n = 6 eyes) and Ptprs−/− retinas (34.5 ± 1.4 ms, n = 6 eyes). (M) Representative flicker ERG raw traces of WT control and Ptprs−/− eyes elicited by 3.162 cd s/m2 flashes at 20 and 30 Hz frequencies. (N) Quantification of step ERG d-wave amplitudes of WT (55.58 ± 6.43 µV n = 6 eyes) and Ptprs−/− (46.40 ± 5.94 µV n = 6 eyes) elicited by a 3-s step of light at 1,000 cd/m2 intensity. (O) Representative step ERG raw traces of WT (n = 6) control and Ptprs−/− (n = 6) eyes elicited by a 3-s step light at 1,000 cd/m2 intensity. Scale bars: 20 µm. Error bars: SEM.
Figure S6.

In situ hybridization of Grik1 and Grm6 and immunostaining of GRM6 upon loss of Fat3 in mouse retina (Related to Fig. 7 ). (A–A″) VSX2 immunostaining (A) after in situ hybridization for Grik1 (A′) and Grm6 (A″) in WT retina. (B–B″) VSX2 immunostaining (B) after in situ hybridization for Grik1 (B′) and Grm6 (B″) in Fat3∆TM/∆TM retina. (C) Quantification of Grik1 RNA mean fluorescence intensity in bipolar cells. WT: 1.00 ± 0.03, n = 4 animals, 16 retinal regions; Fat3∆TM/∆TM: 0.75 ± 0.04, n = 4 animals, 17 retinal regions. Each data point corresponds to a retinal region, color-coded by animal, analyzed using a nested two-tailed test. (D) Quantification of Grm6 RNA mean fluorescence intensity in bipolar cells. WT: 1.00 ± 0.02, n = 4 animals, from 16 retinal regions; Fat3∆TM/∆TM: 0.98 ± 0.02, n = 4 animals, from 17 retinal regions. Each data point corresponds to a retinal region, color-coded by animal, analyzed using a nested two-tailed test. (E) Immunostaining for GRM6 in WT retinas. (F) Immunostaining for GRM6 in Fat3∆TM/∆TM retinas. (E′ and F′) Cone arrestin (ARR3) labels the cone photoreceptors endings in the OPL in E′ and F′. (G) Immunostaining for GRM6 in WT retinas, littermates of Fat3∆ICD-GFP/∆ICD-GFP animals. (H) Immunostaining for GRM6 in Fat3∆ICD-GFP/∆ICD-GFP retinas. Cone arrestin (ARR3) labels the cone photoreceptors endings in the OPL in G′ and H′. (I) Quantification of GRM6 integrated intensity in the OPL. WT controls: 1.00 ± 0.08 (n = 8 animals, 35 retinal regions); Fat3∆TM/∆TM: 1.15 ± 0.09 (n = 9 animals, 41 retinal regions). Each data point corresponds to a retinal region, color-coded by animal, nested two-tailed test. (J) Quantification of GRM6 integrated intensity in the OPL. WT Controls: 1.00 ± 0.09 (n = 5 animals, 21 retinal regions); Fat3∆ICD/∆ICD: 0.86 ± 0.07 (n = 5 animals, 22 retinal regions). Each data point corresponds to a retinal region, color-coded by animal, analyzed using a nested two-tailed test. (K) Flicker ERG amplitude at 30 Hz for WT (18.70 ± 2.05 µV n = 6 eyes) and Ptprs−/− retinas (24.50 ± 2.86 µV, n = 6 eyes). (L) Flicker ERG implicit time at 20 Hz for WT (34.3 ± 0.6 ms, n = 6 eyes) and Ptprs−/− retinas (34.5 ± 1.4 ms, n = 6 eyes). (M) Representative flicker ERG raw traces of WT control and Ptprs−/− eyes elicited by 3.162 cd s/m2 flashes at 20 and 30 Hz frequencies. (N) Quantification of step ERG d-wave amplitudes of WT (55.58 ± 6.43 µV n = 6 eyes) and Ptprs−/− (46.40 ± 5.94 µV n = 6 eyes) elicited by a 3-s step of light at 1,000 cd/m2 intensity. (O) Representative step ERG raw traces of WT (n = 6) control and Ptprs−/− (n = 6) eyes elicited by a 3-s step light at 1,000 cd/m2 intensity. Scale bars: 20 µm. Error bars: SEM.

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