. 2023 Oct 16;10(10):ENEURO.0054-23.2023. doi: 10.1523/ENEURO.0054-23.2023

Table 2.

Summary of statistical analysis

Experiment	Animal number and sex	Test	p-value	t, df	F, dfn, dfd
Figure 2B	P7: N = 6 (3 females and 3 males); P17: N= 4 (2 females and 2 males); 4W: N = 4 (2 females and 2 males); 7W: N = 7 (3 females and 4 males)
tdT⁺ CR cell density, hippocampal fissure		Nested one-way ANOVA	<0.0001		79.85, 3, 59
Figure 2B
tdT⁺ CR cell density, lower molecular layer		Nested one-way ANOVA	0.0095; **<0.0001		178.3, 3, 59
Figure 2B
tdT⁺ granule neurons, suprapyramidal blade		Nested one-way ANOVA	0.0367; **0.0003		6.819, 3, 59
Figure 2B
tdT⁺ granule neurons, infrapyramidal blade		Nested one-way ANOVA	**0.0024		5.054, 3, 59
Figure 2B			P7 vs P17: 0.0491; P7 vs 4W: ***<0.0001
CR cell-targeting specificity		Nested one-way ANOVA	P7 vs 7W: 0.0369; 4W vs 7W: 0.0179		9.611, 3, 59
Extended Data Figure 2-1
tdT⁺ CR cell density, neocortex		Nested one-way ANOVA	<0.0001		148.7, 3, 59
Extended Data Figure 2-3
tdT⁺ granule neurons, suprapyramidal blade, P17		Nested t test	0.1918	t = 1.400, df = 10	1.960, 1, 10
Extended Data Figure 2-3
tdT⁺ granule neurons, suprapyramidal blade, 4W		Nested t test	0.2669	t = 1.176, df = 10	1.382, 1, 10
Extended Data Figure 2-3
tdT⁺ granule neurons, suprapyramidal blade, 7W		Nested t test	0.4237	t = 0.8177, df = 19	0.6686, 1, 19
Extended Data Figure 2-3
tdT⁺ granule neurons, infrapyramidal blade, P17		Nested t test	0.212	t = 1.333, df = 10	1.777, 1, 10
Extended Data Figure 2-3
tdT⁺ granule neurons, infrapyramidal blade, 4W		Nested t test	0.5247	t = 0.6592, df = 10	0.4345, 1, 10
Extended Data Figure 2-3
tdT⁺ granule neurons, infrapyramidal blade, 7W		Nested t test	0.4111	t = 0.8405, df = 19	0.7064, 1, 19
Figure 3B	4W: N = 4 (2 females and 2 males); 7W: N = 7 (2 females and 2 males)
tdT⁺ cells, 4W		Welch's t test	0.1813		t = 1.730, df = 3.025
Figure 3B
tdT⁺ cells, 7W		Welch's t test	0.0424		t = 3.362, df = 3.060
Figure 4C	P14: n = 1 female and 1 male; 5W: n = 1 female and 3 males
CR cell density		Nested t test	<0.0001	t = 11.02, df = 19	121.4, 1, 19
Figure 4C
Granule neuron density		Nested t test	0.2771	t = 1.219, df = 5	1.487, 1, 5
Figure 4D
CR cell density (hippocampal fissure)		Nested one-way ANOVA	0.0194; *0.0013		14.25, 2, 9
Figure 4D
CR cell density (lower molecular layer)	ArchT^ΔNp73-Cre: n = 3 males	Nested one-way ANOVA	0.0015; *0.0003		23.03, 2, 9
Figure 4D	ChETA^ΔNp73-Cre: n = 2 females and 1 male
Granule neuron density (suprapyramidal blade)	hM3Dq^ΔNp73-Cre: n = 1 female and 3 males	Nested one-way ANOVA	0.0017; **<0.0001		14.34, 2, 36
Figure 4D
Granule neuron density (infrapyramidal blade)		Nested one-way ANOVA	*0.0003; **<0.0001		21.16, 2, 36
Figure 4D
CR cell targeting specificity		Nested one-way ANOVA	****<0.0001		28.05, 2, 36
Figure 4E
Variation between hemispheres (ArchT^ΔNp73-Cre)	n = 3 males	Nested t test	0.1259	t = 2.545, df = 2	6.478, 1, 2
Figure 4E
Variation between hemispheres (hM3Dq^ΔNp73-Cre)	n = 1 female and 3 males	Nested t test	0.1308	t = 2.486, df = 2	6.180, 1, 2
Extended Data Figure 4-2D
CR cell density (Layer 1)	ArchT^ΔNp73-Cre: n = 3 males	Nested one-way ANOVA	Not significant		1.863, 2, 17
	ChETA^ΔNp73-Cre: n = 2 females and 1 male
	hM3Dq^ΔNp73-Cre: n = 1 female and 3 males
Figure 5B	1.0 × 10¹¹: N = 5 (1 female and 4 males); 1.0 × 10¹²: N = 4 (1 female and 3 males); 1.0 × 10¹³: N = 5 (4 females and 1 male);
Percentage of mCherry⁺ CRs, hippocampal fissure		Nested one-way ANOVA	0.0013; **<0.0001		261.2, 2, 6
Figure 5B
Percentage of mCherry⁺ CRs, lower molecular layer		Nested one-way ANOVA	0.0016; 0.0001; ***<0.0001		152.8, 2, 6
Figure 5B
Percentage of mCherry⁺ granule neurons		Nested one-way ANOVA	10^11 vs. 10^13: 0.0032 (); 10^12 vs. 10^13: 0.0025 ()		8.613, 2, 39
Extended Data Figure 5-3C	1.0 × 10¹¹ at P14: N = 5 (2 females and 3 males); 1.0 × 10¹² at P14: N = 6 (1 female and 5 males); 1.0 × 10¹³ at P14: N = 3 (2 females and 1 male); 1.0 × 10¹² at 7W: N = 3 (3 females)
Percentage of mCherry⁺ CRs, hippocampal fissure		Nested one-way ANOVA	all not significant		0.8421, 3, 47
Extended Data Figure 5-3C
Percentage of mCherry⁺ CRs, lower molecular layer		Nested one-way ANOVA	all not significant		0.5963, 3, 47
Extended Data Figure 5-3C
Number of mCherry⁺ granule neurons		Nested one-way ANOVA	all not significant		1.443, 3, 8
Extended Data Figure 5-3D	WT: N = 3 (2 females and 1 male)
Number of mCherry⁺ granule neurons	ΔNp73-Cre: N = 5 (4 females and 1 male)	Nested t test	0.0313		5.290, 1, 22
Figure 6B	WT: n = 3 (3 females)
Percentage of mCherry⁺ CRs, hippocampus	ΔNp73-Cre: n = 10 (3 females and 7 males)	Nested t test	<0.0001	t = 11.61, df = 36	134.9, 1, 36
Figure 6C	P0 to >P14: n = 4 (1 female and 3 males)
Percentage of mCherry⁺ granule neurons	P0 to >7W: n = 10 (3 females and 7 males)	Nested t test	0.0506	t = 2.017, df = 39	4.069, 1, 39
Figure 6D	tdTΔNp73-Cre: n = 7 (3 females and 4 males)
Reporter-expressing granule neurons at 7W	AAV-ChR2: n = 10 (3 females and 7 males)	Nested one-way ANOVA	***0.0006 and 0.0007		9.297, 2, 83
Figure 6D	AAV-hM3D: n = 12 (5 females and 7 males)
CR targeting specificity		Nested one-way ANOVA	***0.0003 and 0.0009		9.563, 2, 83
Figure 6E	WT, P14: n = 6 (1 female and 5 males)
Percentage of mCherry⁺ CRs, hippocampal fissure	WT, 7W: n = 3 (3 females)	Two-way ANOVA	Interaction: 0.2082		1.673, 1, 24
Figure 6E	ΔNp73-Cre, P14: n = 4 (1 female and 3 males)
Percentage of mCherry⁺ CRs, lower molecular layer	ΔNp73-Cre, 7W: n = 10 (3 females and 7 males)	Two-way ANOVA	Interaction: 0.9824		0.0004962, 1, 24
Figure 7B
mCherry⁺ CRs at P14, hippocampal fissure	AAV/ChR2: n = 4 (1 female and 3 males)	Nested t test	<0.0001	t = 5.563, df = 19	30.94, 1, 19
Figure 7B	AAV/hM3D: n = 6 (1 female and 5 males)
mCherry⁺ CRs at P14, lower molecular layer		Nested t test	0.0002	t = 4.696, df = 19	22.05, 1, 19
Figure 7D
mCherry⁺ CRs at 7W, hippocampal fissure	AAV/ChR2: n = 10 (3 females and 7 males)	Nested t test	0.0011	t = 3.422, df = 63	11.71, 1, 63
Figure 7D	AAV/hM3D: n = 12 (5 females and 7 males)
mCherry⁺ CRs at 7W, lower molecular layer		Nested t test	0.0072	t = 2.779, df = 63	7.724, 1, 63
Figure 7E	P0 to >P14: n = 6 (1 female and 5 males)
mCherry⁺ CRs at 7W, lower molecular layer	P0 to >7W: n = 12 (5 females and 7 males)	Nested t test	0.0011	t = 3.467, df = 52	12.02, 1, 52
Figure 7F	Wild type, P14: n = 8 (6 females and 2 males)
Total CRs, hippocampal fissure	Wild type, 7W: n = 7 (2 females and 5 males)	Two-way ANOVA	Interaction: 0.4793		0.5136, 1, 29
Figure 7F	ΔNp73-Cre, P14: n = 6 (1 female and 5 males)
Total CRs, lower molecular layer	ΔNp73-Cre, 7W: n = 12 (5 females and 7 males)	Two-way ANOVA	Interaction: 0.8317		0.04601, 1, 29
Extended Data Figure 7-1C
Percentage of mCherry⁺ CRs, hippocampal fissure		Nested t test	0.2784	t = 1.098, df = 42	1.206, 1, 42
Extended Data Figure 7-1C	P0 to >P14: n = 8 (6 females and 2 males)
Percentage of mCherry⁺ CRs, lower molecular layer	P0 to >7W: n = 7 (2 females and 5 males)	No test was performed as there was no variation among replicates
Extended Data Figure 7-1C
mCherry⁺ granule neuron density		No test was performed as there was no variation among replicates
Extended Data Figure 7-2	Female, AAV/ChR2: n = 3
Percentage of mCherry⁺ CRs at 7W, hippocampal fissure	Male, AAV/ChR2: n = 7	Nested one-way ANOVA	**0.0066		4.502, 3, 58
Extended Data Figure 7-2	Female, AAV/hM3D: n = 5
Percentage of mCherry⁺ CRs at 7W, lower molecular layer	Male, AAV/hM3D: n = 7	Nested one-way ANOVA	Not significant		2.638, 3, 61
Figure 8C
Percentage of mCherry⁺ CR cells, hippocampal fissure		Nested t test	<0.0001	t = 10.30, df = 34	106.0, 1, 34
Figure 8C
Percentage of mCherry⁺ CR cells, lower molecular layer	Saline: n = 5 (2 females and 3 males)	Nested t test	<0.0001	t = 10.55, df = 34	111.2, 1, 34
Figure 8C	CNO: n = 7 (3 females and 4 males)
Percentage of total⁺ CR cells, hippocampal fissure		Nested one-way ANOVA	****<0.0001		98.35, 3, 68
Figure 8C
Percentage of total CR cells, lower molecular layer		Nested one-way ANOVA	****<0.0001		114.2, 3, 68
Extended Data Figure 8-1
Percentage of total⁺ CR cells, hippocampal fissure	Saline: n = 5 (5 males)	Nested one-way ANOVA	Not significant		0.7748, 3, 8
Extended Data Figure 8-1	CNO: n = 2 (2 females)
Percentage of total⁺ CR cells, lower molecular layer		Nested one-way ANOVA	Not significant		0.6000, 3, 36

W, Weeks; P, postnatal days; CR, Cajal-Retzius cells; WT, wild type; tdT, tdTomato; df, degree of freedom. dfn, degrees of freedom in the numerator; dfd, degrees of freedom in the denominator.