Extended Data Fig. 8: Efficiency of PKM2 deletion in the SSp and SNc.
a, Upper, Schematic diagram of injection site (modified from Allen Mouse Brain Atlas, online version 1, 2008). Middle, an image showing expression of AAV-hSyn-Cre in NSE-positive neurons. Lower, quantification showing Cre-expressing NSE-positive cells comprise 82% of the total NSE-positive neurons in the SSp (n = 8 mice). b-g, RNAscope analysis of pkm1 (f) and pkm2 (b) mRNA expression or PKM2 immunofluorescence signals (d) in the somata of cortical neurons labelled with the neuron-specific enolase (NSE) antibody in the SSp of Pkm2flox/flox mice injected with AAV-hSyn-Cre unilaterally (right) for one month. The number of pkm2 mRNA spots per soma in the cortical neurons of the SSp with (n = 212 from 3 mice) or without (n = 239 from 3 mice) injection of AAV-hSyn-Cre, p < 0.0001 (c). Quantification of somatic PKM2 fluorescence intensity in the cortical neurons of the SSp with (n = 220 from 3 mice) or without (193 from 3 mice) injection of AAV-hSyn-Cre, p < 0.0001 (e). The number of pkm1 mRNA spots per soma in the cortical neurons of the SSp with (n = 205 from 3 mice) or without (n = 197 from 3 mice) injection of AAV-hSyn-Cre (g). h, Upper, Schematic diagram of injection site (modified from Allen Mouse Brain Atlas, online version 1, 2008). Middle, an image showing expression of AAV-TH-Cre in TH-positive neurons. Lower, quantification showing Cre-expressing TH-positive cells comprise 75% of the total TH-positive neurons in the SNc (n = 8 mice). i-n, RNAscope analysis of pkm1 (m) and pkm2 (i) mRNA expression or PKM2 immunofluorescence signals (k) in the somata of dopaminergic neurons labelled with the tyrosine hydroxylase (TH) antibody in the SNc of Pkm2flox/flox mice injected with AAV-TH-Cre unilaterally (right) for one month. The number of pkm2 mRNA spots per soma in the dopaminergic neurons of the SNc with (n = 218 from 3 mice) or without (n = 205 from 3 mice) injection of AAV-TH-Cre, p < 0.0001 (j). Quantification of somatic PKM2 fluorescence intensity in the dopaminergic neurons of the SNc with (n = 193 from 3 mice) or without (n = 224 from 3 mice) injection of AAV-TH-Cre, p < 0.0001 (l). The number of pkm1 mRNA spots per soma in the dopaminergic neurons of the SNc with (n = 185 from 3 mice) or without (n = 211 from 3 mice) injection of AAV-TH-Cre (n). For the boxplots, the centre line indicates the median, the box limits indicate the first and third quartiles, and the whiskers indicate the data range. Scale bars, 20 μm. Data are presented as mean ± SEM and ***p < 0.001. Student’s two-tailed unpaired t-test (c, e, g, j, l and n).
