Journal: Cell reports

Article Title: Batf3 + DCs and the 4–1BB/4–1BBL axis are required at the effector phase in the tumor microenvironment for PD-1/PD-L1 blockade efficacy

doi: 10.1016/j.celrep.2024.114141

Figure Lengend Snippet: (A) 4–1BB expression on CD8 + SIY + T cells in each condition. Experimental design is shown in , n ≥ 15 per group. (B) Representative histograms. Mean fluorescence intensity (MFI) values are shown inside the graph. (C) Relative 4–1BBL expression, expressed as relative MFI (rMFI = MFI/MFI of FMO) on pDCs, DC1s, and DC2s at day 9 of tumor injection, 1 day after one dose of anti-PD-L1 treatment. Gating strategy described in , n = 16 per group. (D) Percentage of pDCs, DC1s, and DC2s that acquired dsRed (tumor Ag) with and without anti-PD-L1 treatment, n = 16 per group. (E) Expression levels of 4–1BBL on DC1s that did or did not acquire dsRed. One of two independent experiments is shown, n = 8 per group. (F) Number of pDCs, DC1s, and DC2s per gram of tumor that acquired dsRed and express 4–1BBL upon anti-PD-L1 treatment, n = 16 per group. (G) Tumor growth curves of B16·SIY cells injected in WT or 4–1BB KO mice and treated or not with anti-PD-L1, n ≥ 10 per group. (H) Number of CD8 + SIY + T cells per gram of tumor in each group at day 21 after tumor injection, n ≥ 9 per group. (I) Tumor growth curves of B16·SIY cells injected in Rag2 KO mice reconstituted with WT or 4–1BB KO splenocytes and treated or not with anti-PD-L1, n = 13 per group. (J) Tumor growth curves of B16·SIY cells injected in C57BL/6 mice, treated every day with FTY720 from day 7 of tumor injection, and treated with or without anti-PD-L1 mAb in the absence or presence of anti-4–1BBL-blocking mAb, n ≥ 9 per group. (K) Number of CD8 + SIY + T cells per gram of tumor at day 15 after tumor injection. Mice were treated as in (J) but analyzed after 3 doses of mAb treatments. Three independent experiments, n ≥ 17 per group. (L) CD8 + SIY + T cell-to-Foxp3 + T cell ratio in each group of mice treated as in (K), n ≥ 12 per group. (M) Tumor growth curves of MC38.SIY cells injected in C57BL/6 mice and treated as in (J), n ≥ 15 per group. Two independent experiments in all graphs unless stated otherwise. Bar graphs represent the mean values of the indicated data points, and the error bars represent SEM. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. One-way ANOVA with Bonferroni’s post-test (A, C, F, H, K, and L), two-way ANOVA with Bonferroni’s post-test (D and E), and two-way ANOVA with Tukey’s post-test (G, I, J, and M) were used for statistical analysis.

Article Snippet: RAG2 knock-out (KO) mice were purchased from Taconic.

Techniques: Expressing, Fluorescence, Injection, Blocking Assay

Journal: Physiological Reports

Article Title: FGFR regulator Memo1 is dispensable for FGF23 expression by osteoblasts during folic acid‐driven kidney injury

doi: 10.14814/phy2.15650

Figure Lengend Snippet: Primers used for qPCR.

Article Snippet: An osteoblast‐specific Memo1 knock‐out (Memo1 obKO) mouse model was established by crossing mice floxed for exon two of the Memo1 gene (Haenzi et al., ) backcrossed to the C57BL/6J background over at least 10 generations with Col1a1 Cre transgenic mice carrying a Cre recombinase under the control of the 2.3‐kb proximal fragment of the α1(I)‐collagen promoter (Dacquin et al., ; Haenzi et al., ) obtained through Riken BioResource Research Center (Ibaraki, Japan).

Techniques:

Journal: Physiological Reports

Article Title: FGFR regulator Memo1 is dispensable for FGF23 expression by osteoblasts during folic acid‐driven kidney injury

doi: 10.14814/phy2.15650

Figure Lengend Snippet: Validation of Memo obKO mouse model. Femoral Memo1 gene expression (a) and protein abundance (b) were blunted in Memo obKO, whereas Cre expression was detectable (c) in marrow‐free femur of Memo obKO. Memo1 expression was conserved in bone marrow (d), kidney (e) and liver (f) of Memo obKO animals relative to controls. Statistical analyses by Mann–Whitney U test. *, p < 0.05; ns, not significant. N of animals is indicated by the number of scatters. Animals were aged 9 weeks and both sexes at equal ratios in each group.

Article Snippet: An osteoblast‐specific Memo1 knock‐out (Memo1 obKO) mouse model was established by crossing mice floxed for exon two of the Memo1 gene (Haenzi et al., ) backcrossed to the C57BL/6J background over at least 10 generations with Col1a1 Cre transgenic mice carrying a Cre recombinase under the control of the 2.3‐kb proximal fragment of the α1(I)‐collagen promoter (Dacquin et al., ; Haenzi et al., ) obtained through Riken BioResource Research Center (Ibaraki, Japan).

Techniques: Biomarker Discovery, Gene Expression, Quantitative Proteomics, Expressing, MANN-WHITNEY

Journal: Physiological Reports

Article Title: FGFR regulator Memo1 is dispensable for FGF23 expression by osteoblasts during folic acid‐driven kidney injury

doi: 10.14814/phy2.15650

Figure Lengend Snippet: Three‐dimensional reconstructions of bone sites investigated by micro‐CT. Structure was not affected by loss of Memo1 in osteoblasts at any site, and 1 representative image per anatomical site and genotype is shown.

Article Snippet: An osteoblast‐specific Memo1 knock‐out (Memo1 obKO) mouse model was established by crossing mice floxed for exon two of the Memo1 gene (Haenzi et al., ) backcrossed to the C57BL/6J background over at least 10 generations with Col1a1 Cre transgenic mice carrying a Cre recombinase under the control of the 2.3‐kb proximal fragment of the α1(I)‐collagen promoter (Dacquin et al., ; Haenzi et al., ) obtained through Riken BioResource Research Center (Ibaraki, Japan).

Techniques: Micro-CT

Journal: Physiological Reports

Article Title: FGFR regulator Memo1 is dispensable for FGF23 expression by osteoblasts during folic acid‐driven kidney injury

doi: 10.14814/phy2.15650

Figure Lengend Snippet: Femoral μCT analysis of Memo1 obKO females ( n = 4).

Article Snippet: An osteoblast‐specific Memo1 knock‐out (Memo1 obKO) mouse model was established by crossing mice floxed for exon two of the Memo1 gene (Haenzi et al., ) backcrossed to the C57BL/6J background over at least 10 generations with Col1a1 Cre transgenic mice carrying a Cre recombinase under the control of the 2.3‐kb proximal fragment of the α1(I)‐collagen promoter (Dacquin et al., ; Haenzi et al., ) obtained through Riken BioResource Research Center (Ibaraki, Japan).

Techniques: Micro-CT, Control

Journal: Physiological Reports

Article Title: FGFR regulator Memo1 is dispensable for FGF23 expression by osteoblasts during folic acid‐driven kidney injury

doi: 10.14814/phy2.15650

Figure Lengend Snippet: Tibia μCT analysis of Memo1 obKO males ( n = 4).

Article Snippet: An osteoblast‐specific Memo1 knock‐out (Memo1 obKO) mouse model was established by crossing mice floxed for exon two of the Memo1 gene (Haenzi et al., ) backcrossed to the C57BL/6J background over at least 10 generations with Col1a1 Cre transgenic mice carrying a Cre recombinase under the control of the 2.3‐kb proximal fragment of the α1(I)‐collagen promoter (Dacquin et al., ; Haenzi et al., ) obtained through Riken BioResource Research Center (Ibaraki, Japan).

Techniques: Micro-CT, Control

Journal: Physiological Reports

Article Title: FGFR regulator Memo1 is dispensable for FGF23 expression by osteoblasts during folic acid‐driven kidney injury

doi: 10.14814/phy2.15650

Figure Lengend Snippet: Tibia μCT analysis of Memo1 obKO females ( n = 4).

Article Snippet: An osteoblast‐specific Memo1 knock‐out (Memo1 obKO) mouse model was established by crossing mice floxed for exon two of the Memo1 gene (Haenzi et al., ) backcrossed to the C57BL/6J background over at least 10 generations with Col1a1 Cre transgenic mice carrying a Cre recombinase under the control of the 2.3‐kb proximal fragment of the α1(I)‐collagen promoter (Dacquin et al., ; Haenzi et al., ) obtained through Riken BioResource Research Center (Ibaraki, Japan).

Techniques: Micro-CT, Control

Journal: Physiological Reports

Article Title: FGFR regulator Memo1 is dispensable for FGF23 expression by osteoblasts during folic acid‐driven kidney injury

doi: 10.14814/phy2.15650

Figure Lengend Snippet: Vertebral μCT analysis of Memo1 obKO males ( n = 4).

Article Snippet: An osteoblast‐specific Memo1 knock‐out (Memo1 obKO) mouse model was established by crossing mice floxed for exon two of the Memo1 gene (Haenzi et al., ) backcrossed to the C57BL/6J background over at least 10 generations with Col1a1 Cre transgenic mice carrying a Cre recombinase under the control of the 2.3‐kb proximal fragment of the α1(I)‐collagen promoter (Dacquin et al., ; Haenzi et al., ) obtained through Riken BioResource Research Center (Ibaraki, Japan).

Techniques: Micro-CT, Control

Journal: Physiological Reports

Article Title: FGFR regulator Memo1 is dispensable for FGF23 expression by osteoblasts during folic acid‐driven kidney injury

doi: 10.14814/phy2.15650

Figure Lengend Snippet: Vertebral μCT analysis of Memo1 obKO females ( n = 4).

Article Snippet: An osteoblast‐specific Memo1 knock‐out (Memo1 obKO) mouse model was established by crossing mice floxed for exon two of the Memo1 gene (Haenzi et al., ) backcrossed to the C57BL/6J background over at least 10 generations with Col1a1 Cre transgenic mice carrying a Cre recombinase under the control of the 2.3‐kb proximal fragment of the α1(I)‐collagen promoter (Dacquin et al., ; Haenzi et al., ) obtained through Riken BioResource Research Center (Ibaraki, Japan).

Techniques: Micro-CT, Control

Journal: Physiological Reports

Article Title: FGFR regulator Memo1 is dispensable for FGF23 expression by osteoblasts during folic acid‐driven kidney injury

doi: 10.14814/phy2.15650

Figure Lengend Snippet: Serum parameters of Memo1 obKO ( n = 4 per genotype and sex).

Article Snippet: An osteoblast‐specific Memo1 knock‐out (Memo1 obKO) mouse model was established by crossing mice floxed for exon two of the Memo1 gene (Haenzi et al., ) backcrossed to the C57BL/6J background over at least 10 generations with Col1a1 Cre transgenic mice carrying a Cre recombinase under the control of the 2.3‐kb proximal fragment of the α1(I)‐collagen promoter (Dacquin et al., ; Haenzi et al., ) obtained through Riken BioResource Research Center (Ibaraki, Japan).

Techniques: Control

Journal: Physiological Reports

Article Title: FGFR regulator Memo1 is dispensable for FGF23 expression by osteoblasts during folic acid‐driven kidney injury

doi: 10.14814/phy2.15650

Figure Lengend Snippet: Femoral μCT analysis of Memo1 obKO males ( n = 4).

Article Snippet: An osteoblast‐specific Memo1 knock‐out (Memo1 obKO) mouse model was established by crossing mice floxed for exon two of the Memo1 gene (Haenzi et al., ) backcrossed to the C57BL/6J background over at least 10 generations with Col1a1 Cre transgenic mice carrying a Cre recombinase under the control of the 2.3‐kb proximal fragment of the α1(I)‐collagen promoter (Dacquin et al., ; Haenzi et al., ) obtained through Riken BioResource Research Center (Ibaraki, Japan).

Techniques: Micro-CT, Control

Journal: PLoS ONE

Article Title: Neuropsychological Deficits in Mice Depleted of the Schizophrenia Susceptibility Gene CSMD1

doi: 10.1371/journal.pone.0079501

Figure Lengend Snippet: (A) Schematic representation of the KO-strategy. A 1 kb genomic region (white lines) of exon1/intron1 was replaced with a selection cassette (grey box). (B) Expression of Csmd1 mRNA measured by QPCR in an adult mouse tissue panel. Csmd1 is predominantly expressed in brain tissues as compared to peripheral tissues. The highest expression level was identified in areas of the cortex. (C) Depletion of Csmd1 mRNA in the cortex was documented by two exon-exon specific QPCR assays. Transcription of exon 1–2 was depleted, while about 20% residual expression could be observed when amplifying exon 32–33. KO mice lacked a protein band of expected size (389 KDa, arrow), as demonstrated by immunoblotting. Signals of lower molecular weight are indicated (a and b). (D) Mapping of RNA-seq reads to the Csmd1 locus. RNA sequencing of cortex is shown for 4wild-type (green) and 4 Csmd1 KO (red) mice (transcript scale: 0–150 reads). Coverage signals of modified nucleosomes (H3K4me3, H3K4me1 and H3K27Ac) and polymerase-2 binding profiles are shown for the mouse cortex. The 1 kb deleted sequence of Csmd1 is highlighted in yellow (upper panel) and blue (lower panel). No RNA reads were mapped to the deleted genomic region in the KO mice. Abbreviations: Cx, cortex; VCx, visual cortex; FCx, frontal cortex; Hipp, hippocampus; Hyp, hypothalamus; Ob, olfactory bulb; Cer, cerebellum; Visc. Fat, visceral fat.

Article Snippet: A DNA sequence-verified repository Csmd1 gene knock-out (KO) mouse model (clone TF0137, Taconic, Denmark) was generated with embryonic stem cells derived from 129SvEvBrd mice, in which a 1,070 bp genomic sequences of the exon 1– intron 1 junction was replaced with a LacZ/Neo selection cassette .

Techniques: Selection, Expressing, Western Blot, Molecular Weight, RNA Sequencing Assay, Modification, Binding Assay, Sequencing

Journal: PLoS ONE

Article Title: Neuropsychological Deficits in Mice Depleted of the Schizophrenia Susceptibility Gene CSMD1

doi: 10.1371/journal.pone.0079501

Figure Lengend Snippet: (A) Heat map of RNA-seq expression levels, as measured by rpkm, mapping to individual exons and introns of Csmd1 . The arrow indicates the orientation of exons and introns in their increasing number. (B) Analysis of differentially expressed RNAs. Significantly up-regulated and down-regulated transcripts are represented by red dots. Y-axis indicates the log2 (fold change).

Article Snippet: A DNA sequence-verified repository Csmd1 gene knock-out (KO) mouse model (clone TF0137, Taconic, Denmark) was generated with embryonic stem cells derived from 129SvEvBrd mice, in which a 1,070 bp genomic sequences of the exon 1– intron 1 junction was replaced with a LacZ/Neo selection cassette .

Techniques: RNA Sequencing Assay, Expressing

Journal: PLoS ONE

Article Title: Neuropsychological Deficits in Mice Depleted of the Schizophrenia Susceptibility Gene CSMD1

doi: 10.1371/journal.pone.0079501

Figure Lengend Snippet: (A) Map of RNA sequencing reads aligned to the promoter region of Csmd1. Data from individual Csmd1 KO (N = 4; red lines) and WT (N = 4; green lines) mice are shown. The novel pas-lncRNA RNA (black thick line) is expressed antisense to the Csmd1 promoter sequence (transcript scale: 0–30 reads). (B) Heat map representing the relative expression level of pas-lncRNA and Csmd1 mRNA in peripheral and CNS tissues of Csmd1 KO and WT mice, respectively. Expression values of each transcript are calculated relative to their respective expression level in cortex of WT mice. pas-lncRNA expression was induced in the CNS but not in peripheral tissues of Csmd1 KO mice. Fold change values (FC) and statistical significances are listed for each tissue. (C) Co-regulated expression of Csmd1 and pas-lncRNA in the developing cortex and cerebellum ( Csmd1 : pas-lncRNA expression correlation coefficient, cortex: r 2 = 0.92). Y-axis represents relative expression level of RNA. X-axis indicates the postnatal day. Abbreviations: asterisk, t-test P -value<0.05; n.s.; not significant.

Article Snippet: A DNA sequence-verified repository Csmd1 gene knock-out (KO) mouse model (clone TF0137, Taconic, Denmark) was generated with embryonic stem cells derived from 129SvEvBrd mice, in which a 1,070 bp genomic sequences of the exon 1– intron 1 junction was replaced with a LacZ/Neo selection cassette .

Techniques: RNA Sequencing Assay, Sequencing, Expressing

Journal: PLoS ONE

Article Title: Neuropsychological Deficits in Mice Depleted of the Schizophrenia Susceptibility Gene CSMD1

doi: 10.1371/journal.pone.0079501

Figure Lengend Snippet: (A) Analysis of total time spent on open arms demonstrate significant less time for KO mice (N = 13) as compared to WT mice (N = 8). (B) Compiled tracks from all mice show that Csmd1 KO mice avoid entering open arms, as opposed to WT mice traveling over the entire test arena. Abbreviations: asterisk, statistically significant ( P -value<0.05); s, seconds.

Article Snippet: A DNA sequence-verified repository Csmd1 gene knock-out (KO) mouse model (clone TF0137, Taconic, Denmark) was generated with embryonic stem cells derived from 129SvEvBrd mice, in which a 1,070 bp genomic sequences of the exon 1– intron 1 junction was replaced with a LacZ/Neo selection cassette .

Techniques:

Journal: PLoS ONE

Article Title: Neuropsychological Deficits in Mice Depleted of the Schizophrenia Susceptibility Gene CSMD1

doi: 10.1371/journal.pone.0079501

Figure Lengend Snippet: (A) Startle responses of Csmd1 KO mice in response to acoustic stimuli in the range of 80–120 dB, as compared to WT mice. The startle baseline was similar in male and female WT mice. A marginal increase in startle responses could be observed for higher acoustic stimuli in both genders of Csmd1 KO mice, reaching statistical significance when analyzing all mice together (genotype-group interaction P -value<0.05). (B) There was no difference in amplitude response between KO and WT mice in the degree of inhibition after pre-pulse inhibition. (C) Tail suspension test demonstrated longer accumulated time immobile for KO mice as compared WT mice (P-value<0.05). Borderline statistical significance was observed for male Csmd1 KO mice as compared to WT mice (P-value = 0.1).

Article Snippet: A DNA sequence-verified repository Csmd1 gene knock-out (KO) mouse model (clone TF0137, Taconic, Denmark) was generated with embryonic stem cells derived from 129SvEvBrd mice, in which a 1,070 bp genomic sequences of the exon 1– intron 1 junction was replaced with a LacZ/Neo selection cassette .

Techniques: Inhibition

Journal: PLoS ONE

Article Title: Neuropsychological Deficits in Mice Depleted of the Schizophrenia Susceptibility Gene CSMD1

doi: 10.1371/journal.pone.0079501

Figure Lengend Snippet: (A and B) Object contacts in the first and second trial of exposure to novel and familiar objects demonstrated increased contact counts for Csmd1 KO mice (P-value<0.05). (B and C) Discrimination and preference analysis demonstrated no effect of Csmd1 on object recognition. Abbreviation: n.s., not significant; asterisk, statistically significant ( P -value<0.05).

Article Snippet: A DNA sequence-verified repository Csmd1 gene knock-out (KO) mouse model (clone TF0137, Taconic, Denmark) was generated with embryonic stem cells derived from 129SvEvBrd mice, in which a 1,070 bp genomic sequences of the exon 1– intron 1 junction was replaced with a LacZ/Neo selection cassette .

Techniques: