Table 1 Statistics for the results.

Fig. 1		Time point	Sample size	Test	P value	t, df, or F (Dfn, Dfd)
B	Total-caspase8	24 h	n = 6–7	Unpaired t test, two tailed	0.363	t = 0.9487, df = 11
	43kd cleaved-caspase8	24 h	n = 6–7	Unpaired t test, two tailed	0.002	t = 4.172, df = 11
	18kd cleaved-caspase 8	24 h	n = 6–7	Unpaired t test, two tailed	0.000	t = 5.624, df = 11
C	Caspase 8 activity	24 h	n = 7–8	Unpaired t test, two tailed	<0.0001	t = 6.76, df = 13
E	RIPK1	Sham/3 h/6 h/24 h		One-way anova	0.9953	F (3, 16) = 0.02219
		Sham vs. 3 h	n = 3–7	Dunnett test	0.9979
		Sham vs. 6 h	n = 3–7	Dunnett test	0.9987
		Sham vs. 24 h	n = 7	Dunnett test	0.9993
F	RIPK3	Sham/3 h/6 h/24 h		One-way anova	0.0121	F (3, 16) = 5.033
		Sham vs. 3 h	n = 3–7	Dunnett test	0.0334
		Sham vs. 6 h	n = 3–7	Dunnett test	0.0482
G		Sham vs. 24 h	n = 7	Dunnett test	0.0129
	MLKL	Sham/3 h/6 h/24 h		One-way anova	0.0004	F (3, 17) = 10.55
		Sham vs. 3 h	n = 4–8	Dunnett test	0.0391
		Sham vs. 6 h	n = 3–8	Dunnett test	0.2926
		Sham vs. 24 h	n = 6–8	Dunnett test	0.0001
I	RIPK1 expression in CD31 (WT)	24 h	n = 7	Unpaired t test, two tailed	0.027	t = 2.512 df = 12
	RIPK1 expression in CD11b	24 h	n = 5–6	Unpaired t test, two tailed	0.022	t = 2.76, df = 9
	RIPK1 expression in neuron	24 h	n = 5–6	Unpaired t test, two tailed	0.010	t = 3.28, df = 9
J	RIPK3 expression in CD31	24 h	n = 6	Unpaired t test, two tailed	0.009	t = 3.224, df = 10
	RIPK3 expression in CD11b	24 h	n = 5	Unpaired t test, two tailed	0.083	t = 1.982, df = 8
	RIPK3 expression in neuron	24 h	n = 5	Unpaired t test, two tailed	0.370	t = 0.95, df = 8
K	MLKL epxression in CD31_	24 h	n = 6	Unpaired t test, two tailed	0.924	t = 0.09827, df = 10
	MLKL expression in CD11b	24 h	n = 6	Unpaired t test, two tailed	0.568	t = 0.5903, df = 10
	MLKL expression in neuron	24 h	n = 5	Unpaired t test, two tailed	0.030	t = 2.636, df = 8

Fig. 2		Time point	Sample size	Test	P value	t, df/F(DFn,DFd)
B	RIPK1 in triton fraction	Sham 3 h 24 h		One-way anova	0.270	F (2, 6) = 1.642
		Sham vs 3 h	n = 3	Dunnett test	0.2736
		Sham vs 24 h	n = 3	Dunnett test	0.2725
	RIPK1 in urea fraction	Sham 3 h 24 h		One-way anova	0.0071	F (2, 6) = 12.60
		3 h	n = 3	Dunnett test	0.2786
		24 h	n = 3	Dunnett test	0.0048
C	RIPK3 in trition fraction	Sham 3 h 24 h	n = 3	One-way anova	0.0315	F (2, 6) = 6.503
		3 h	n = 3	Dunnett test	0.0874
		24 h	n = 3	Dunnett test	0.0221
D	MLKL in triton fraction	Sham 3 h 24 h	n = 3	One-way anova	0.1161	F (2, 6) = 3.149
		3 h	n = 3	Dunnett test	0.3572
		24 h	n = 3	Dunnett test	0.0794
	MLKL in urea fraction	Sham 3 h 24 h	n = 3	One-way anova	0.019	F (2, 6) = 8.244
		3 h	n = 3	Dunnett test	0.187
		24 h	n = 3	Dunnett test	0.0119
F	p-RIPK3 in CD11b	3 h	n = 3	Unpaired t test, two tailed	0.011	t = 4.536, df = 4
	p-RIPK3 in neuron	3 h	n = 4	Unpaired t test, two tailed	<0.0001	t = 10.25, df = 6
	p-MLKL in neuron	3 h	n = 3	Unpaired t test, two tailed	0.041	t = 2.973, df = 4
H	p-RIPK3 in CD31	24 h	n = 3	Unpaired t test, two tailed	0.011	t = 4.485, df = 4
	p-RIPK3 in CD11b	24 h	n = 3	Unpaired t test, two tailed	0.044	t = 2.910, df = 4
	p-RIPK3 in neuron	24 h	n = 4	Unpaired t test, two tailed	0.045	t = 2.520, df = 6
	p-MLKL in neuron	24 h	n = 4	Unpaired t test, two tailed	0.023	t = 3.041, df = 6
K	p-RIPK3 3D culture	24 h	n = 6	Unpaired t test, two tailed	0.003	t = 3.828, df = 10
	RIPK3 3D culture	24 h	n = 3	Unpaired t test, two tailed	0.046	t = 2.852, df = 4

Fig. 3		Comparison	Sample size	Test	Column factor		Time x column factor		Time		Subject
B	RIPK3KO wire grip		n = 19–21	Two-way RM ANOVA	0.019	F (1, 38) = 6.033	0.140	F (8, 304) = 1.549	<0.0001	F (4.535, 172.3) = 85.86	<0.0001	F (38, 304) = 2.681
C	RIPK3KO rotarod	Pre WT vs RIPK3KO	n = 9–10	Unpaired t test, two tailed	0.545	t = 0.6177, df = 17
		Post WT vs RIPK3KO	n = 9–11	Two-way RM ANOVA	0.017	F (1, 17) = 6.961	0.876	F (3, 51) = 0.2291	<0.0001	F (1.892, 32.16) = 26.30	0.279	F [17, 51] = 1.226
D	RIPK3KO mwm	Hidden platform	n = 9–12	Two-way RM ANOVA	0.028	F (1, 19) = 5.667	0.279	F (6, 114) = 1.266	<0.0001	F (4.105, 77.99) = 8.564	<0.0001	F (19, 114) = 5.452
		Visible platform	n = 9–10	Two-way RM ANOVA	0.253	F (1, 17) = 1.399	0.000	F [1, 17] = 20.55	0.632	F [1, 17] = 0.2385	0.164	F [17, 17] = 1.623
E	RIPK3KO probe test	WT pre vs WT post	n = 9	Unpaired t test, two tailed	0.005	t = 3.220, df = 16
		RIPK3−/− pre vs post	n = 12	Unpaired t test, two tailed	0.680	t = 0.4176, df = 22
F	RIPK3KO NORT	WT pre old vs novel	n = 10	Paired t test	0.001	t = 4.852, df = 9
		WT post old vs novel	n = 10	Paired t test	0.615	t = 0.5203, df = 9
		RIPK3KO pre old vs novel	n = 9	Paired t test	0.001	t = 5.464, df = 8
		RIPK3KO post old vs novel	n = 9	Paired t test	0.033	t = 2.565, df = 8
G	MLKLKO wire grip	Wire grip	n = 14–16	Two-way RM ANOVA	0.314	F (1, 28) = 1.051	0.237	F (6, 152) = 1.354	<0.0001	F (4.133, 104.7) = 32.89
H	MLKL KO rotarod	WT pre vs MLKL pre	n = 12–13	Unpaired t test, two tailed	0.969	t = 0.03919, df = 23
		WT post vs MLKL post	n = 12–13	Two-way RM ANOVA	0.700	F (1, 23) = 0.1523	0.972	F (3, 69) = 0.07760	0.131	F (1.597, 36.73) = 2.238	<0.0001	F [23, 69] = 3.342
I	MLKLKO MWM	Hidden platform	n = 14–15	Two-way RM ANOVA	0.321	F (1, 27) = 1.020	0.079	F (6, 162) = 1.931	<0.0001	F (3.562, 96.19) = 7.131	<0.0001	F (27, 162) = 7.589
		Visible platform	n = 14–15	Two-way RM ANOVA	0.003	F (1, 27) = 11.08	0.253	F (1, 27) = 1.366	0.573	F [1, 27] = 0.3257	0.144	F [27, 27] = 1.514
J	MLKLKO probe test	WT pre vs WT post	n = 14–16	Unpaired t test, two tailed	0.001	t = 3.948, df = 29
		MLKL KO pre vs post CCI	n = 14–17	Unpaired t test, two tailed	0.049	t = 2.062, df = 27
		WT post CCI vs MLKL post CCI	n = 14–18	Unpaired t test, two tailed	0.408	t = 0.8410, df = 27

Fig. 4		Region	Sample size	Test	p value	t, df
B	RIPK3KO vs WT PI	CA1	n = 8	Unpaired t test, two tailed	0.725	t = 0.3591, df = 14
		CA3	n = 8	Unpaired t test, two tailed	0.562	t = 0.5943, df = 14
		DG	n = 8	Unpaired t test, two tailed	0.090	t = 1.820, df = 14
		Cortex	n = 8	Unpaired t test, two tailed	0.721	t = 0.3641, df = 14
D	FJB + cell in WT vs RIPK3KO	CA1	n = 6	Unpaired t test, two tailed	0.213	t = 1.331, df = 10
		CA3	n = 6	Unpaired t test, two tailed	0.775	t = 0.2943, df = 10
		DG	n = 6	Unpaired t test, two tailed	0.187	t = 1.418, df = 10
		Cortex middle	n = 6	Unpaired t test, two tailed	0.504	t = 0.6931, df = 10
		Cortex lateral	n = 6	Unpaired t test, two tailed	0.979	t = 0.02650, df = 10
F	WT vs RIPK3−/− lesion volume		n = 9–10	Unpaired t test, two tailed	0.632	t = 0.4872, df = 17
H	WT vs MLKL−/− lesion volume		n = 14–16	Unpaired t test, two tailed	0.436	t = 0.7903, df = 28

Fig. 5					Ipsilateral		Contralateral				WT con vs ipsi	KO con vs ipsi
		Time point	Sample size	Test	p value	t, df	p value	t, df			p value	p value
B	RIPK3 vs WT evansblue	24 h	n = 7	Unpaired t test, two tailed	0.044	t = 2.250, df = 12	0.601	t = 0.5375, df = 12	0.022		2.522E-07	8.856E-05
D	WT vs MLKL/-/ evans blue	24 h	n = 6	Unpaired t test, two tailed	0.027	t = 2.580, df = 10	0.260	t = 1.195, df = 10	0.021		4.142E-06	6.657E-05
E	WT vs RIPK3-/- edema	24 h	n = 4–5	Unpaired t test, two tailed	0.162	t = 1.565, df = 7	0.484	t = 0.7387, df = 7	0.157	t = 1.583, df = 7	6.696E-06	1.066E-04
F	WT vs MLKL edema	24 h	n = 6	Unpaired t test, two tailed	0.095	t = 1.847, df = 10	0.026	t = 2.613, df = 10	0.586	t = 0.5625, df = 10	3.087E-05	3.678E-02

Fig. 6		Comparison	Sample size	Test	p value	t, df
B	CD11b 48 h	Sham WT vs RIPK3KO	n = 4	Unpaired t test, two tailed	0.529	t = 0.6678, df = 6
		CCI WT vs RIPK3KO	n = 6	Unpaired t test, two tailed	0.016	t = 2.879, df = 10
C	Microglia 48 h	Sham WT vs RIPK3KO	n = 4	Unpaired t test, two tailed	0.604	t = 0.5477, df = 6
		CCI WT vs RIPK3KO	n = 6	Unpaired t test, two tailed	0.027	t = 2.595, df = 10
D	Macrophage 48 h	Sham WT vs RIPK3KO	n = 4	Unpaired t test, two tailed	0.910	t = 0.1177, df = 6
		CCI WT vs RIPK3KO	n = 6	Unpaired t test, two tailed	0.013	t = 3.028, df = 10
E	Neutrophil 48 h	Sham WT vs RIPK3KO	n = 4	Unpaired t test, two tailed	0.529	t = 0.6675, df = 6
		CCI WT vs RIPK3KO	n = 6	Unpaired t test, two tailed	0.221	t = 1.307, df = 10
F	Lymphocytes 48 h	Sham WT vs RIPK3KO	n = 4	Unpaired t test, two tailed	0.581	t = 0.5833, df = 6
		CCI WT vs RIPK3KO	n = 6	Unpaired t test, two tailed	0.443	t = 0.7990, df = 10
G	IL1b elisa brain tissue	Sham WT vs RIPK3KO	n = 5–6	Unpaired t test, two tailed	0.007	t = 3.451, df = 9
		CCI WT vs RIPK3KO	n = 6–9	Unpaired t test, two tailed	0.002	t = 3.978, df = 13
I	CD11b + 3 w	WT vs RIPK3KO	n = 5–6	Unpaired t test, two tailed	0.042	t = 2.376, df = 9
J	Microglia 3 w	WT vs RIPK3KO	n = 5–6	Unpaired t test, two tailed	0.037	t = 2.450, df = 9
K	Macrophage 3 w	WT vs RIPK3KO	n = 5–6	Unpaired t test, two tailed	0.628	t = 0.5014, df = 9
L	Lymphocytes 3 w	WT vs RIPK3KO	n = 5–6	Unpaired t test, two tailed	0.971	t = 0.03764, df = 9

Fig. 7		Time point	Sample size	Test	p value	t, df
C	Neurons	24 h	n = 3	Unpaired t test, two tailed	0.011	t = 4.436, df = 4
E	WT	24 h	n = 5	Unpaired t test, two tailed	0.006	t = 5.409, df = 4
	RIPK3−/−	24 h	n = 4	Unpaired t test, two tailed	0.448	t = 0.8714, df = 3
	MLKL	24 h	n = 3	Unpaired t test, two tailed	<0.0001	t = 105.7, df = 2

Supplement Fig. 2
B	p-RIPK1 sham vs CCI	24 h	n = 6	Unpaired t test, two tailed	0.525	t = 0.6587,df = 10
	RIPK1 sham vs CCI	24 h	n = 6	Unpaired t test, two tailed	0.368	t = 0.9419,df = 10
C	p-MLKL sham vs CCI	24 h	n = 6	Unpaired t test, two tailed	0.306	t = 1.079,df = 10
	MLKL sham vs CCI	24 h	n = 6	Unpaired t test, two tailed	0.586	t = 0.5626, df = 10

Supplement Fig. 3		Sample size	Test	p value		Time x column factor		t, df
A	RIPK3KO vs WT baseline mwm	n = 9–12	Two-way RM ANOVA	0.468	F (1, 19) = 0.5478	0.966	F (6, 114) = 0.2314
B	MLKLKO vs WT baseline mwm	n = 15–16	Two-way RM ANOVA	0.823	F (1, 29) = 0.05105	0.909	F (6, 174) = 0.3505
C	RIPK3KO vs WT platform crossing	n = 9–12	Unpaired t test, two tailed	0.282				t = 1.106, df = 19
D	MLKLKO vs WT platform crossing	n = 15–16	Unpaired t test, two tailed	0.820				t = 0.2296, df = 27
E	RIPK3KO vs WT post CCI swim speed	n = 9–12	Two-way RM ANOVA	0.207	F (1, 19) = 1.709	P = 0.0921	F (6, 114) = 1.869
F	MLKLKO vs WT post CCI swim speed	n = 14–15	Two-way RM ANOVA	0.267	F (1, 27) = 1.285	0.766	F (6, 162) = 0.5546

Table 1 Statistics for the results.

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