![Focal knockdown of spinal total upstream frameshift 1 (UPF1) expression relieves spinal nerve ligation-induced allodynia-like behaviors in rats. (A) Representative immunoblotting and statistical analysis (normalized to glyceraldehyde 3-phosphate dehydrogenase [GAPDH]) of total UPF1 and phosphorylated UPF1 contents in the dorsal horn of naïve rats on Day 4 after total UPF1 small-interfering RNA (total UPF1 small-interfering RNA; 5 μg; 10 μl; once daily for 4 days) or missense small-interfering RNA (5 μg, 10 μl) injection. **P < 0.01 versus naïve group. Experimental groups: three. Each group: five rats. Total UPF1, one-way ANOVA, F(2,12) = 15.25, P < 0.001. Phosphorylated UPF1, one-way ANOVA, F(2,12) = 8.61, P = 0.005. (B) Rotarod test measured among naïve, missense small-interfering RNA (5 μg; 10 μl; once daily for 4 days), and total UPF1 small-interfering RNA groups (5 μg; 10 μl; once daily for 4 days). Experimental groups: three. Each group: six rats. Two-way ANOVA, group, F(2,15) = 1.49, P = 0.258; time, F(4,60) = 0.79, P = 0.538; interaction, F(8,60) = 0.53, P = 0.829. (C and D) The paw withdrawal threshold measured of sham operation (Sham) and spinal nerve ligation rats in response to intrathecal administration of missense small-interfering RNA (5 μg; 10 μl; daily from Days 3 to 6 after operation) or total UPF1 small-interfering RNA groups (5 μg; 10 μl; daily from Days 3 to 6 after operation). **P < 0.01 versus spinal nerve ligation group. Experimental groups: three (C) and three (D). Each group: six rats. (C) Two-way ANOVA, group, F(2,15) = 0.45, P = 0.647; time, F(4,60) = 0.90, P = 0.471; interaction, F(8,60) = 0.65, P = 0.736. (D) Two-way ANOVA, group, F(2,15) = 0.48, P = 0.025; time, F(4,60) = 106.6, P < 0.001; interaction, F(8,60) = 8.18, P < 0.001. (E) Representative Western blot and statistical analyses (normalized to GAPDH) total UPF1 and phosphorylated UPF1 in the ipsilateral dorsal horn on Day 7 of spinal nerve ligation rats in response to intrathecal administration of total UPF1 small-interfering RNA (spinal nerve ligation Day 7 + total UPF1 small-interfering RNA, 5 μg, 10 μl, daily from Days 3 to 6 after operation) or missense small-interfering RNA (spinal nerve ligation Day 7 + missense small-interfering RNA, 5 μg; 10 μl; daily from Days 3 to 6 after operation). **P < 0.05 versus sham group on Day 7. ##P < 0.01 versus spinal nerve ligation group on Day 7. Experimental groups: four. Each group: five rats. Total UPF1, one-way ANOVA, F(3,16) = 10.46, P < 0.001. Phosphorylated UPF1, one-way ANOVA, F(3,16) = 14.69, P < 0.001.](https://asa2.silverchair-cdn.com/asa2/content_public/journal/anesthesiology/138/6/10.1097_aln.0000000000004550/1/15ff02.jpeg?Expires=1716434494&Signature=PrJRaNYQU~o~DWRXpkF17OTJFKApp8eBInbXn1Za1-4SVA~JG1GJFsOhuOTmWsww7d7VccSFUdB5F1EFWP7GP7c8UCPEU~TsNySiyXg-rXpbcfXjmyb5m22P9Xmr-ZU9W8jWLA~Yd2L7F-xKkmOnVjB2DMoqWXhBUYEWofvwnJ7PGfGENurEsou0-GBc2zmCmBHXRvgTaMeNVWo4uyYOulz-P5iVxgi2oOwWjralKEpts~y8mp8I3VyPVmPMJ1OiNiiPtw4g9EFhcYYKwyeHXnucdFDQMzfSoZfr-LZpMss3OCvcEymFmJPBMzvkk1lhHXQo-je4BN2ayL9nBBUfog__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Focal knockdown of spinal total upstream frameshift 1 (UPF1) expression relieves spinal nerve ligation-induced allodynia-like behaviors in rats. (A) Representative immunoblotting and statistical analysis (normalized to glyceraldehyde 3-phosphate dehydrogenase [GAPDH]) of total UPF1 and phosphorylated UPF1 contents in the dorsal horn of naïve rats on Day 4 after total UPF1 small-interfering RNA (total UPF1 small-interfering RNA; 5 μg; 10 μl; once daily for 4 days) or missense small-interfering RNA (5 μg, 10 μl) injection. **P < 0.01 versus naïve group. Experimental groups: three. Each group: five rats. Total UPF1, one-way ANOVA, F(2,12) = 15.25, P < 0.001. Phosphorylated UPF1, one-way ANOVA, F(2,12) = 8.61, P = 0.005. (B) Rotarod test measured among naïve, missense small-interfering RNA (5 μg; 10 μl; once daily for 4 days), and total UPF1 small-interfering RNA groups (5 μg; 10 μl; once daily for 4 days). Experimental groups: three. Each group: six rats. Two-way ANOVA, group, F(2,15) = 1.49, P = 0.258; time, F(4,60) = 0.79, P = 0.538; interaction, F(8,60) = 0.53, P = 0.829. (C and D) The paw withdrawal threshold measured of sham operation (Sham) and spinal nerve ligation rats in response to intrathecal administration of missense small-interfering RNA (5 μg; 10 μl; daily from Days 3 to 6 after operation) or total UPF1 small-interfering RNA groups (5 μg; 10 μl; daily from Days 3 to 6 after operation). **P < 0.01 versus spinal nerve ligation group. Experimental groups: three (C) and three (D). Each group: six rats. (C) Two-way ANOVA, group, F(2,15) = 0.45, P = 0.647; time, F(4,60) = 0.90, P = 0.471; interaction, F(8,60) = 0.65, P = 0.736. (D) Two-way ANOVA, group, F(2,15) = 0.48, P = 0.025; time, F(4,60) = 106.6, P < 0.001; interaction, F(8,60) = 8.18, P < 0.001. (E) Representative Western blot and statistical analyses (normalized to GAPDH) total UPF1 and phosphorylated UPF1 in the ipsilateral dorsal horn on Day 7 of spinal nerve ligation rats in response to intrathecal administration of total UPF1 small-interfering RNA (spinal nerve ligation Day 7 + total UPF1 small-interfering RNA, 5 μg, 10 μl, daily from Days 3 to 6 after operation) or missense small-interfering RNA (spinal nerve ligation Day 7 + missense small-interfering RNA, 5 μg; 10 μl; daily from Days 3 to 6 after operation). **P < 0.05 versus sham group on Day 7. ##P < 0.01 versus spinal nerve ligation group on Day 7. Experimental groups: four. Each group: five rats. Total UPF1, one-way ANOVA, F(3,16) = 10.46, P < 0.001. Phosphorylated UPF1, one-way ANOVA, F(3,16) = 14.69, P < 0.001.
