Fig. 3: Loss of ATF4 impaired heat nociception, and re-expression of ATF4 rescued heat nociception.
From: ATF4 selectively regulates heat nociception and contributes to kinesin-mediated TRPM3 trafficking
a–j The behaviours of WT, Atf4+/− and rescued mice were evaluated by the von Frey (a), dynamic (b), tape (c), tail clip (d), pinprick (e), Hargreaves (f), tail-flick (g), hot plate (h), evaporative cooling (i) and rotarod (j) tests. n = 10 mice in WT, n = 7 mice in Atf4+/− and n = 6 mice in rescue group. F(2,20) = 12.11, P = 0.0004 in WT vs. Atf4+/−, P = 0.0050 in Atf4+/− vs. rescue in f. F(2,20) = 12.59, P = 0.0008 in WT vs. Atf4+/−, P = 0.0009 in Atf4+/− vs. rescue in 48 °C; F(2,20) = 20.67, P < 0.0001 in WT vs. Atf4+/−, P = 0.0002 in Atf4+/− vs. rescue in 50 °C; F(2,20) = 8.924, P = 0.0024 in WT vs. Atf4+/−, P = 0.0076 in Atf4+/− vs. rescue in 52 °C in g. F(2,20) = 7.097, P = 0.0159 in WT vs. Atf4+/−, P = 0.0066 in Atf4+/− vs. rescue in 50 °C; F(2,20) = 5.678, P = 0.0135 in WT vs. Atf4+/−, P = 0.0375 in Atf4+/− vs. rescue in 52 °C; F(2,20) = 21.31, P < 0.0001 in WT vs. Atf4+/−, P < 0.0001 in Atf4+/− vs. rescue in 55 °C in h. k, l Knockout and rescue of ATF4 impaired and rescued SNL-induced heat hyperalgesia (k) but not mechanical allodynia (l), respectively. n = 10 mice in WT, n = 9 mice in Atf4+/− and n = 9 mice in rescue group. F(2,25) = 36; P = 0.0038 in WT vs. Atf4+/−, P = 0.0098 in Atf4+/− vs. rescue in day 0; P = 0.0002 in WT vs. Atf4+/−, P < 0.0001 in Atf4+/− vs. rescue in day 4; P = 0.0145 in WT vs. Atf4+/−, P = 0.0170 in Atf4+/− vs. rescue in day 7; P = 0.0105 in WT vs. Atf4+/−, P = 0.0034 in Atf4+/− vs. rescue in day 10; P = 0.0174 in WT vs. Atf4+/−, P = 0.0087 in Atf4+/− vs. rescue in day 14 in k. F(2,25) = 1.112; P = 0.0171 in WT vs. Atf4+/−, P = 0.0074 in Atf4+/− vs. rescue in day 14 in l. m, n Knockout and rescue of ATF4 impaired and rescued CFA-induced heat hyperalgesia (m) but not mechanical allodynia (n), respectively. n = 10 mice in WT, n = 9 mice in Atf4+/− and n = 9 mice in rescue group. F(2,25) = 108.5; P = 0.0002 in WT vs. Atf4+/−, P = 0.0005 in Atf4+/− vs. rescue in day 0; P = 0.0045 in WT vs. Atf4+/−, P = 0.0086 in Atf4+/− vs. rescue in day 1; P = 0.0001 in WT vs. Atf4+/−, P = 0.0017 in Atf4+/− vs. rescue in day 3; P = 0.0006 in WT vs. Atf4+/−, P < 0.0001 in Atf4+/− vs. rescue in day 5. o, p Spontaneous pain: total duration (o) or number (p) of nocifensive behaviour (paw licking or flinching within 2 min) in response to intraplantar injection of vehicle, PS (2.5 nmol/paw) or CIM0216 (2.5 nmol/paw) into WT, Atf4+/− and rescued mice. n = 10 mice per group. F(2,27) = 6.375, P = 0.0368 in WT vs. Atf4+/−, P = 0.0056 in Atf4+/− vs. rescue in PS; F(2,27) = 9.174, P = 0.0027 in WT vs. Atf4+/−, P = 0.0026 in Atf4+/− vs. rescue in CIM0216 in o. F(2,27) = 5.976, P = 0.0201 in WT vs. Atf4+/−, P = 0.0123 in Atf4+/− vs. rescue in PS; F(2,27) = 7.569, P = 0.0033 in WT vs. Atf4+/−, P = 0.0138 in Atf4+/− vs. rescue in CIM0216 in p. q Spontaneous pain: total duration of nocifensive behaviour (in 5 min) in response to intrathecal injection of vehicle, PS (1.25 nmol/paw) or CIM0216 (1.25 nmol/paw) into WT, Atf4+/− and rescued mice. n = 10 mice per group. F(2,27) = 5.849, P = 0.0194 in WT vs. Atf4+/−, P = 0.0146 in Atf4+/− vs. rescue in PS; F(2,27) = 5.986, P = 0.0127 in WT vs. Atf4+/−, P = 0.0192 in Atf4+/− vs. rescue in CIM0216. a–j, o–q, One-way ANOVA followed by Tukey’s multiple comparisons test. k–n Two-way ANOVA followed by Bonferroni’s multiple comparisons test. *P < 0.05, **P < 0.01, n.s. means not significant. The errors bars indicate the SEMs.
