Extended Data Fig. 2: LPAR4 does not impact cell growth in the absence of stress. | Nature Cell Biology

Extended Data Fig. 2: LPAR4 does not impact cell growth in the absence of stress.

From: Pancreatic cancer cells upregulate LPAR4 in response to isolation stress to promote an ECM-enriched niche and support tumour initiation

Extended Data Fig. 2

a, Immunohistochemistry staining of LPAR4 in a PDAC tissue array consisted of 15 PDAC samples and 4 samples of normal pancreas. Scale bar is 50 μM. Representative images showing normal pancreas (n = 4 biological samples), low-LPAR4 PDAC (n = 9 biological samples), and high-LPAR4 PDAC (n = 6 biological samples). Bar graph shows the relative percentage of LPAR4-low and LPAR4-high patients in the PDAC tissue array. b, Quantitative RT-PCR confirming the ectopic expression and stable knockdown of LPAR4 in 2 pancreatic cancer cell lines (Colo-357, MIA PaCa-2) and 2 patient-derived cancer cells (79E, 34E). Data were shown as mean ± s.d. (n = 4 independent experiments for Colo-357 and 79E cells with LPAR4 stable knockdown, n = 3 for Colo-357, 79E, and MiaPaCa2 cells with LPAR4 ectopic expression, n = 3 for 34E cells with LPAR4 stable knockdown, and n = 4 for 34E with LPAR4 ectopic expression). c, Non-invasive Bioluminescence images showing tumors formed at day 10 for Colo-357+sh-CTRL+ luciferase or Colo-357+sh-R4.1+luciferase of various number implanted in the pancreas of nu/nu mice. The right panel showing the luminescence intensity in a blue-to-red spectrum. d, f, Trypan blue exclusion assay showing the relative viable cell number of cells with or without LPAR4 expression manipulation grown in 10% serum and 2D at day 4 and day 7. e, Tumor growth rates for 1 million Colo-357 cells with or without LPAR4 expression manipulation in a subcutaneous tumor model. Data were presented as mean ± s.d. for n = 8 independent samples for each group. g, Quantitative RT-PCR confirming the knockdown of LPAR1 in Colo-357 cells. h, Effects of LPAR1 knockdown using siRNA on 2D or 3D (methylcellulose sphere forming) cell growth. Data were presented as mean ± s.d. for n = 3 independent experiments (d, f, g, and h). Statistical analyses were performed using two tailed unpaired one sample t-test (b, g, and h) and one-way ANOVA (d–f). Source numerical data are available in source data.

Source data

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