rnascope probe-mm-tgfb2-c2  (Advanced Cell Diagnostics Inc)

Journal: Nature metabolism

Article Title: Diet-dependent regulation of TGFβ impairs reparative innate immune responses after demyelination

doi: 10.1038/s42255-021-00341-7

Figure Lengend Snippet: a, Quantification of the increase in mRNA expression of Apoe, Abca1 and Abcg1 from 2 to 7dpi lesions from Cx3cr1creERT,wt:Tgfbr2fí/f, (Tgfbr2 KO) or Cx3cr1wt,wt:Tgfbr2fl/fl (Tgfbr2 Control) mice fed a WD (two-tailed Welch’s t-test). b,d, Quantification of demyelination (b) and IBA1+ (d) volume at 14dpi in Tgfbr2 control and KO mice treated with tamoxifen 14 days before lesion induction and fed WD (two-tailed Welch’s t-test). c,f, Quantification of demyelination (c) and IBA1+ (f) volume at 14dpi in Tgfbr2 control and KO mice treated with tamoxifen 14 days before lesion induction and fed CD (two-tailed Welch’s t-test). e, Images of corpus callosum lesions at 14 dpi. Scale bar: 200 μm. g,h Quantification of the number of myelin- (g) and crystal-loaded (h) IBA1+ cells per mm2 of lesion (two-tailed Welch’s t-test). i, Images of the demyelinated lesion in the corpus callosum at 14dpi showing myelin- and crystal-loaded IBA1+ cells. White arrows point to myelin- and crystal-loaded IBA1+ cells. Scale bar, 20 μm. j, The site of action of the conditional genetic knock-out and mode of action of GS in the TGFβ pathway. GS, galunisertib. k,l, Quantification of demyelination (k) and IBA1+ (l) volume at 14dpi (one-way Brown-Forsythe and Welch ANOVA tests with multiple comparisons corrected by Dunnett T3 test). m, Images of corpus callosum lesions at 14 dpi. Scale bar, 200 μm. n,o, Quantification of the number of myelin- (n) and crystal-loaded (o) IBA1+ cells per mm2 of lesion (two-tailed Welch’s t-test). p, Images of the demyelinated lesion in the corpus callosum at 14dpi exemplifying myelin- and crystal-loaded IBA1+ cells. White arrows point to myelin- and crystal-loaded IBA1+ cells. Scale bar, 20μm. P-values below 0.1 and n numbers are indicated in the figure; each dot represents one mouse. Solid lines in the graphs indicate the mean.

Article Snippet: First, 16-μm-thick brain sections were hybridized with the respective mRNA probes: RNAscope Probe-Mm-Tgfb1, Cat. No. 407751; RNAscope Probe-Mm-Tgfb2-C2, Cat. No. 406181-C2; RNAscope Probe-Mm-Abca1, Cat. No. 522251; RNAscope Probe-Mm-Abcg1-C2, Cat. No. 422221-C2; and RNAscope Probe-Mm-Tgfbr2, Cat. No.;406241, all from Advanced Cell Diagnostics.

Techniques: Expressing, Control, Two Tailed Test, Knock-Out

Journal: Nature metabolism

Article Title: Diet-dependent regulation of TGFβ impairs reparative innate immune responses after demyelination

doi: 10.1038/s42255-021-00341-7

Figure Lengend Snippet: a, Fold change in Tgfbr2 expression in the demyelinated lesions of Tgfbr2 KO and control mice (two-tailed Welch’s t-test). b, Quantification of the number of Tgfbr2’ microglia in the unlesioned brains of Tgfbr2 KO and control mice (two-tailed Welch’s t test). c, Images of the unlesioned brain of Tgfbr2 KO and control mice demonstrating the accumulation of Tgfbr2 RNA particles within IBA1+ microglia. Scale bar: 20μm. d, Quantification of the ratio of TGFβR2 signal over GAPDH signal in microglia isolated from Tgfbr2 KO and control mice (two-tailed t-test). e, Example images of the Western blots used for quantification of TGFβR2 levels of isolated microglia from Tgfbr2 KO and control mice. TGFβR2 levels were normalized to GAPDH. f,g, Quantification of the demyelination (f) and IBA1+ (g) volume at 4 dpi in Tgfbr2 KO and control mice fed WD. h,i, Quantification of the demyelination (h) and IBA1+ (i) volume at 4dpi in Tgfbr2 control and KO mice fed CD. j, Quantification of Tgfbl and Tgfb2 expression in the unlesioned brain of young (3-months old) and old (12-months old) mice by RT-qPCR (two-tailed Welch’s t-test). k,l, Quantification of demyelination k) and IBA1+ (l) volume at 14 dpi in young, old and old+GS mice (one-way Brown-Forsythe and Welch ANOVA tests with multiple comparisons corrected by Dunnett T3 test). m, Images of corpus callosum lesions of old and old+GS mice at 14dpi. Scale bar: 200μm. n,o, Quantification of the number of myelin- (n) and crystal- (o) loaded IBA1+ cells per mm2 of lesion (two-tailed Welch’s t-test). p, Images of the demyelinated lesion in the corpus callosum at 14dpi exemplifying myelin- and crystal-loaded IBA1+ cells in old and old+GS mice. Scale bar: 20μm. N numbers are indicated in the figure; each dot represents one mouse. Solid lines in the graphs indicate the mean. CD: control diet, WD: Western diet GS: Galunisertib, dpi: days post injection.

Article Snippet: First, 16-μm-thick brain sections were hybridized with the respective mRNA probes: RNAscope Probe-Mm-Tgfb1, Cat. No. 407751; RNAscope Probe-Mm-Tgfb2-C2, Cat. No. 406181-C2; RNAscope Probe-Mm-Abca1, Cat. No. 522251; RNAscope Probe-Mm-Abcg1-C2, Cat. No. 422221-C2; and RNAscope Probe-Mm-Tgfbr2, Cat. No.;406241, all from Advanced Cell Diagnostics.

Techniques: Blocking Assay, Expressing, Control, Two Tailed Test, Isolation, Western Blot, Quantitative RT-PCR, Injection